Lettuce variety NUN 09161 LTL

ABSTRACT

The disclosure relates to lettuce variety NUN 09161 LTL as well as seeds and plants and heads or leaves thereof. NUN 09161 LTL is a multileaf green lettuce compact variety comprising resistance to Downy Mildew (Bremia lactucae) Isolates B1:16-37EU and Nasonovia ribisnigri Biotype Nr:0.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to U.S. Provisional Application No.62/939,457, filed on Nov. 22, 2019, which is hereby incorporated byreference in its entirety.

FIELD OF THE DISCLOSURE

The disclosure relates to the field of plant breeding and, morespecifically, to the development of lettuce variety NUN 09161 LTL. Thedisclosure further relates to vegetative reproductions of lettucevariety NUN 09161 LTL, methods for tissue culture of lettuce variety NUN09161 LTL, and to phenotypic variants of lettuce variety NUN 09161 LTL.The disclosure also relates to progeny of lettuce variety NUN 09161 LTLand the hybrid varieties obtained by crossing lettuce variety NUN 09161LTL as a parent line with plants of other varieties or parent lines.

BACKGROUND OF THE DISCLOSURE

The goal of vegetable breeding is to combine various desirable traits ina single variety. Such desirable traits may include greater yield,resistance to diseases, insects or other pests, tolerance to heat anddrought, better agronomic quality, higher nutritional value, enhancedgrowth rate, and improved shelf life.

The development of commercial lettuce cultivars or varieties requiresthe crossing of lettuce plants, and the evaluation of the crosses.Pedigree breeding and recurrent selection are examples of breedingmethods used to develop cultivars from breeding populations. Breedingprograms combine desirable traits from two or more varieties or variousbroad-based sources into breeding pools from which cultivars aredeveloped by selfing and selection of desired phenotypes. The newcultivars are crossed with other varieties and the inbred lines orhybrids from these crosses are evaluated to determine which havecommercial potential.

All cultivated forms of lettuce belong to the highly polymorphic speciesLactuca sativa that is grown for its edible head and leaves. Lactucasativa is in the Asteraceae (Compositae) family. Lettuce is related tochicory, sunflower, aster, dandelion, artichoke, and chrysanthemum. L.sativa is one of about 300 species in the genus Lactuca. There are manytypes of lettuce, and new types are constantly in development. Types oflettuce include Cutting/Leaf, Iceberg/Crisphead, Cos or Romaine,Batavian, Salinas Group, Latin, Butterhead, Great Lakes Group, Eastern(Ithaca) Group, Bibb, Vanguard Group, Multileaf, or Stem lettuce.Lettuce is typically consumed fresh and occasionally as a cookedvegetable. It is popularly used in salads, wraps, and sandwiches.

Fresh lettuce is available in the United States year-round although thegreatest supply is from May through October. For planting purposes, thelettuce season is typically divided into three categories, early, midand late, with the coastal areas planting from January to August, andthe desert regions planting from August to December. California andArizona are the two largest producers of lettuce in the United States.

Changes in lifestyle primarily due to increasing health awarenessresults to growing demand for healthy convenience food. Supermarkets,restaurants, catering firms, and convenience stores are constantlylooking for more colorful garnishing for sandwiches, wraps, andready-to-eat snacks such as salads. The changing food and consumertrends present opportunities for breeding companies to develop newvarieties with specific shapes of leaves, specific average size ofleaves, prominent color, glossiness, taste, and a wide variety oftexture. Other breeding objectives include disease or pest resistance,yield, prolonged shelf life, and suitability to climatic requirements.

SUMMARY OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure provides for lettuce variety NUN 09161 LTL, productsthereof, and methods of using the same. NUN 09161 LTL is a multileafgreen compact lettuce variety and is suitable for growing in the openfield.

In another aspect, the plant of lettuce variety NUN 09161 LTL, a partthereof, or a progeny thereof comprises resistance to Downy Mildew(Bremia lactucae) Isolates B1:16-37EU and Nasonovia ribisnigri BiotypeNr:0, measured according to UPOV standards described in TG/13/11.

In another aspect, the disclosure provides for a hybrid lettuce varietyNUN 09161 LTL. The disclosure also provides for a progeny of lettucevariety NUN 09161 LTL. In another aspect, the disclosure provides aplant or a progeny retaining all or all but one, two, or three of the“distinguishing characteristics” of lettuce variety NUN 09161 LTL, orall but one, two, or three of the “morphological and physiologicalcharacteristics” of lettuce variety NUN 09161 LTL and methods ofproducing that plant or progeny.

In one aspect, the disclosure provides a plant or a progeny having allof the physiological and morphological characteristics of lettucevariety NUN 09161 LTL, when grown under the same environmentalconditions. In another aspect, the plant or progeny has all or all butone, two, or three of the physiological and morphologicalcharacteristics of lettuce variety NUN 09161 LTL when grown under thesame environmental conditions and e.g., evaluated at significance levelsof 1%, 5%, or 10% significance (which can be expressed as a p-value) forquantitative characteristics and determined by type or degree fornon-quantitative characteristics, wherein a representative sample ofseed of variety NUN 09161 LTL has been deposited under Accession NumberNCIMB 43998. In another aspect, the plant or progeny has all or all butone, two, or three of the physiological and morphologicalcharacteristics as listed in Table 1-3 of lettuce variety NUN 09161 LTLwhen measured under the same environmental conditions and e.g.,evaluated at significance levels of 1%, 5%, or 10% significance (whichcan also be expressed as a p value) for quantitative characteristics anddetermined by type or degree for non-quantitative characteristics.

In another aspect, the plant of lettuce variety NUN 09161 LTL, or partthereof, or a progeny thereof has 7, 8, or more or all of the followingdistinguishing characteristics when compared to Reference Variety asshown in Table 4: 1) larger plant spread of frame leaves; 2) shorterplant; 3) longer base of head to apex (core length); 4) larger base ofhead (core width); 5) fewer mature leaf divisions; 6) yellowish huegreen color of mature leaves; 6) glossier mature leaves; 7) strongerundulation of margin of mature leaves; and 8) medium depth of incisionsof margin of mature leaves, when determined at the 5% significance levelfor numerical characteristics and determined by type or degree fornon-numerical characteristics when grown under the same environmentalconditions.

In another aspect, the disclosure provides a seed of lettuce variety NUN09161 LTL, wherein a representative sample of said seed has beendeposited under Accession Number NCIMB 43998. The disclosure alsoprovides for a plurality of seeds of lettuce variety NUN 09161 LTL. Thelettuce seed of variety NUN 09161 LTL may be provided as an essentiallyhomogeneous population of lettuce seed. The population of seed oflettuce variety NUN 09161 LTL may be particularly defined as anessentially free from other seed. The seed population may be grown intoplants to provide an essentially homogeneous population of lettuceplants described herein.

The disclosure also provides a plant grown from a seed of lettucevariety NUN 09161 LTL and a plant part thereof.

The disclosure further provides a lettuce head and/or a lettuce leafproduced on a plant grown from a seed of lettuce variety NUN 09161 LTL.

The disclosure furthermore provides a seed growing or grown on a plantof lettuce variety NUN 09161 LTL (e.g., produced after pollination ofthe flower of lettuce variety NUN 09161 LTL).

In another aspect, the disclosure provides for a plant part obtainedfrom lettuce variety NUN 09161 LTL, wherein said plant part is: a leaf,a part of a leaf, a head, a part of a head, a fruit, a part of a fruit,pollen, an ovule, a cell, a petiole, a shoot or a part thereof, a stemor a part thereof, a root or a part thereof, a root tip, a cutting, aseed, a part of a seed, seed coat or another maternal tissue which ispart of a seed grown on said variety, a hypocotyl, a cotyledon, apistil, an anther, or a flower or a part thereof. Heads and leaves areparticularly important plant parts. Such plant parts may be suitable forsexual reproduction, vegetative reproduction, or a tissue culture. Inanother aspect, the plant part obtained from variety NUN 09161 LTL is acell, optionally a cell in a cell or tissue culture. That cell may begrown into a plant of lettuce variety NUN 09161 LTL.

In another aspect, the disclosure provides for an inbred variety of NUN09161 LTL.

In another aspect, the disclosure provides a cell culture of lettucevariety NUN 09161 LTL and a plant regenerated from lettuce variety NUN09161 LTL, wherein the plant has all of the characteristics of lettucevariety NUN 09161 LTL when grown under the same environmentalconditions, as well as methods for culturing and regenerating lettucevariety NUN 09161 LTL. Alternatively, a regenerated plant may have onecharacteristic that is different from lettuce variety NUN 09161 LTL.

The disclosure also provides a vegetatively propagated plant of varietyNUN 09161 LTL having all or all but one, two, or three of themorphological and physiological characteristics of lettuce variety NUN09161 LTL when grown under the same environmental conditions as well asmethods for vegetatively propagating lettuce variety NUN 09161 LTL.

In another aspect, the disclosure provides a method of producing alettuce plant comprising crossing lettuce variety NUN 09161 LTL withitself or another lettuce variety and selecting a progeny lettuce plantfrom said crossing or selfing.

The disclosure also provides a method of producing a lettuce plantderived from lettuce variety NUN 09161 LTL.

In further aspect, the disclosure provides a method of producing hybridlettuce seed comprising crossing a first parent lettuce plant with asecond parent lettuce plant and harvesting the resultant hybrid lettuceseed, wherein said first parent lettuce plant or second parent lettuceplant is lettuce variety NUN 09161 LTL. Also provided is a hybridlettuce seed produced from crossing a first parent lettuce plant with asecond parent lettuce plant and harvesting the resultant hybrid lettuceseed, wherein said first parent lettuce plant or second parent lettuceplant is lettuce variety NUN 09161 LTL. Moreover, a hybrid lettuce plantgrown from the hybrid lettuce seed is provided.

In another aspect, the disclosure provides a method of introducing asingle locus conversion into a plant of variety NUN 09161 LTL, wherein arepresentative sample of said seed has been deposited under AccessionNumber NCIMB 43998, wherein the plant otherwise retains all of thephysiological and morphological characteristics of lettuce variety NUN09161 LTL and further comprises the single locus conversion.

In yet another aspect, the disclosure provides a method of introducing adesired trait into lettuce variety NUN 09161 LTL, said method comprisestransforming the plant of variety NUN 09161 LTL with a transgene thatconfers the desired trait, wherein the transformed plant otherwise hasall of the physiological and morphological characteristics of lettucevariety NUN 09161 LTL and contains the desired trait.

The disclosure also provides a method of producing a modified lettuceplant with a desired trait, wherein the method comprises mutating alettuce plant or plant part of lettuce variety NUN 09161 LTL, wherein arepresentative sample of said seed has been deposited under AccessionNumber NCIMB 43998, and wherein the mutated plant otherwise retains allof the physiological and morphological characteristics of lettucevariety NUN 09161 LTL and contains the desired trait.

In one aspect, the single locus conversion or desired trait is yield,size, storage properties, color, taste, enhanced nutritional quality,post-harvest quality, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, or modified proteinmetabolism, or the mutation occurs in any of the following genes:Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL, PPO.

In another aspect, the disclosure provides a container comprising theplant, plant part, or seed of lettuce variety NUN 09161 LTL.

Also provided is a food, a feed, or a processed product comprising theplant part of lettuce variety NUN 09161 LTL, wherein the plant part is aleaf or part thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the mature plants of lettuce variety NUN 09161 LTL.

FIG. 2 shows the mature plants of the Reference Variety.

FIG. 3 shows the mature leaves of lettuce variety NUN 09161 LTL.

FIG. 4 shows the mature leaves comparison of lettuce variety NUN 09161LTL and the Reference Variety.

FIG. 5 shows the cross-section of mature leaves of lettuce variety NUN09161 LTL.

FIG. 6 shows the cross-section of mature leaves of the ReferenceVariety.

DEFINITIONS

“Lettuce” refers herein to plants of the species Lactuca sativa L. Themost commonly eaten parts of a lettuce plant are the head or a leaf. Thehead comprises a core and leaves, which may be divided in inner andouter leaves.

“Cultivated lettuce” refers to plants of Lactuca sativa (e.g.,varieties, breeding lines or cultivars of the species L. sativa as wellas crossbreds thereof, or crossbreds with other Lactuca sativa species,or even with other Lactuca species), cultivated by humans and havinggood agronomic characteristics.

“Leaf lettuce” or “cutting lettuce” refers to a type of lettuce havingvery loose leaves that does not form a head.

The terms “lettuce plant designated NUN 09161 LTL,” “NUN 09161 LTL,”“inbred NUN 09161,” “09161 LTL,” “lettuce 09161” or “Multigreen 161” areused interchangeably herein and refer to a lettuce plant of variety NUN09161 LTL, representative sample of said seed has been deposited underAccession Number NCIMB 43998.

“Plant” includes the whole plant or any parts or derivatives thereof,preferably having the same genetic makeup as the plant from which it isobtained.

“Plant part” includes any part of a plant, such as a plant organ (e.g.,harvested or non-harvested fruits), a plant cell, a plant protoplast, aplant cell tissue culture or a tissue culture from which a whole plantcan be regenerated, a plant cell that is intact in a plant, a clone, amicropropagation, plant callus, a plant cell clump, a plant transplant,a vegetative propagation, a fruit, a harvested fruit, a part of a fruit,a leaf, a part of a leaf, pollen, an ovule, an embryo, a petiole, ashoot or a part thereof, a stem or a part thereof, a root or a partthereof, a root tip, a cutting, a seed, a part of a seed, a hypocotyl, acotyledon, a scion, a graft, a stock, a rootstock, a pistil, an anther,or a flower. Seed can be mature or immature. Pollen or ovules may beviable or non-viable. Also, any developmental stage is included, such asseedlings, cuttings prior or after rooting, mature plants or leaves.Alternatively, a plant part may also include a plant seed whichcomprises one or two sets of chromosomes derived from the parent plant(e.g., from lettuce variety NUN 09161 LTL). An F1 progeny produced fromself-pollination of the inbred variety NUN 09161 LTL will thus comprisetwo sets of chromosomes derived from lettuce variety NUN 09161 LTL,while an F1 progeny derived from cross-fertilization of NUN 09161 LTLwill comprise only one set of chromosomes from lettuce variety NUN 09161LTL and the other set of chromosomes from the other parent.

A “seed of lettuce variety NUN 09161 LTL” refers to a lettuce seed whichcan be grown into a plant of lettuce variety NUN 09161 LTL, wherein arepresentative sample of viable seeds of lettuce variety NUN 09161 LTLhas been deposited under Accession Number NCIMB 43998. A seed can be inany stage of maturity, for example, a mature, viable seed, or animmature non-viable seed. A seed comprises an embryo and maternaltissues.

An “embryo of lettuce variety NUN 09161 LTL” refers to an embryo aspresent in a seed of lettuce variety NUN 09161 LTL, a representativesample of said seed of NUN 09161 LTL having been deposited underAccession Number NCIMB 43998.

A “seed grown on lettuce variety NUN 09161 LTL” refers to a seed grownon a mature plant of variety NUN 09161 LTL or inside a fruit of lettucevariety NUN 09161 LTL. The “seed grown on lettuce variety NUN 09161 LTL”contains tissues and DNA of the maternal parent, lettuce variety NUN09161 LTL. The “seed grown on lettuce variety NUN 09161 LTL” contains anF1 embryo. When said seed is planted, it grows into a first generationprogeny plant of variety NUN 09161 LTL. Since NUN 09161 LTL is an inbredvariety and thus highly homozygous, the set of chromosomes inherited bythe first generation is predictable.

An “essentially homogeneous population of lettuce seed” is a populationof seeds where at least 77%, 98%, 99% or more of the total population ofseed are seed of lettuce variety NUN 09161 LTL.

An “essentially homogeneous population of lettuce plants” is apopulation of plants where at least 97%, 97%, 99% or more of the totalpopulation of plants are plants of lettuce variety NUN 09161 LTL.

The phrase “essentially free from other seed” refers to a population ofseeds where less than 3%, 2%, 1%, or even less, of the total populationof seed is seed that is not a lettuce seed or, in another option, lessthan 3%, 2%, 1%, or less, of the total population of seed is seed thatis not seed of lettuce variety NUN 09161 LTL.

“Cotyledon” refers to one of the first leaves of the embryo of a seedplant.

“Head” as used herein refers to lettuce heads, i.e., the plant withoutthe root system, for example, substantially all harvested leaves.Encompassed are immature leaves (e.g., “baby leaf”) and mature leaves.

The “base” of a plant is the part of a lettuce plant where the leavesare attached to the root system of the plant.

“Core length” of the internal lettuce stem is measured from the base ofthe cut and trimmed head to the tip of the stem.

“Core Length to Head Diameter Ratio (CLHD Ratio)” refers to the meancore length/head diameter ratio. It is calculated by dividing the meancore length with the mean head diameter. This is an indication of thehead shape and of the ability of a lettuce plant to reduce the amount ofsurface which is on or close to the ground.

“Head weight” refers to the mean weight of saleable lettuce head, cut,and trimmed to market specifications.

“Head diameter” refers to the mean diameter of the cut and trimmed head,sliced vertically, and measured at the widest point perpendicular to thestem.

“Head height” refers to the mean height of the cut and trimmed head,sliced vertically, and measured from the base of the cut stem to theleaf tip.

“Harvested plant material” refers herein to plant parts (e.g., leaves orheads detached from the whole plant) which have been collected forfurther storage and/or further use.

“Harvested seeds” refers to seeds harvested from a line or variety,e.g., produced after self-fertilization or cross-fertilization andcollected.

“First water date” refers to the date the seed first receives adequatemoisture to germinate. This can and often does equal the planting date.

“Maturity date” refers to the stage when the plants are of full size oroptimum weight, in marketable form or shape to be of commercial oreconomic value. This is also the time when measuring parameters of“mature” leaves.

“Yield” means the total weight of all lettuce heads or leaves harvestedper hectare of a particular line or variety. It is understood that“yield” expressed as weight of all lettuce heads or leaves harvested perhectare can be obtained by multiplying the number of plants per hectaretimes the “yield per plant.”

“Marketable yield” means the total weight of all marketable lettuceheads or leaves harvested per hectare of a particular line or variety,e.g., lettuce heads or leaves suitable for being sold for freshconsumption, having good color, glossiness size and texture and no orvery low levels of deficiencies. A “marketable lettuce head or leaf” isa head or leaf that has commercial value.

“USDA descriptors” are the plant variety descriptors described forlettuce in the “Objective description of Variety—Lettuce (Lactuca sativaL.),” as published by U.S. Department of Agriculture, AgriculturalMarketing Service, Science and Technology, Plant Variety ProtectionOffice (June 2015, revised December 2018), and which can be downloadedfrom the world-wide web at ams.usda.gov/underservices/plant-variety-protection/pvpo-c-forms under lettuce. “Non-USDAdescriptors” are other descriptors suitable for describing lettuce.

“UPOV descriptors” are the plant variety descriptors described forlettuce in the “Guidelines for the Conduct of Tests for Distinctness,Uniformity and Stability,” TG/13/11 (Geneva 2006, last updated 2019 Jun.14), as published by UPOV (International Union for the Protection of NewVarieties and Plants) and which can be downloaded from the world-wideweb at upov.int/under edocs/tgdocs/en/tg013.pdf, which is herebyincorporated by reference in its entirety. Likewise, “UPOV methods” todetermine specific parameters for the characterization of lettuce aredescribed at upov.int.

“RHS” or “RHS color” refers to the color chart of the RoyalHorticultural Society (UK), which publishes a botanical color chartquantitatively identifying colors according to a defined numberingsystem. The chart may be purchased from Royal Horticulture SocietyEnterprise Ltd RHS Garden; Wisley, Woking; Surrey GU236QB, UK, e.g., theRHS color chart 2007.

“Calibration Manual: DUS Test for Lettuce” refers to the calibrationbook for lettuce which provides guidance for describing a lettucevariety, as published by Naktuinbow (Netherlands) and NationalAgriculture and Food Research Organization (NARO Japan) (Dec. 26, 2016)and based on the UPOV Guideline TG/13/10 Rev. 2

“Reference Variety” refers herein to variety NUN 09060 LTL, a varietyfrom Nunhems B.V. with commercial name Multigreen 60, which has beenplanted in a trial together with lettuce variety NUN 09161 LTL. Thecharacteristics of lettuce variety NUN 09161 LTL were compared with thecharacteristics of the Reference Variety as shown in Tables 2 and 3. Thedistinguishing characteristics between lettuce variety NUN 09161 LTL andthe Reference Variety are shown in Table 4.

A plant having “all the physiological and morphological characteristics”of a referred-to-plant means a plant showing the physiological andmorphological characteristics of the referred-to-plant when grown underthe same environmental conditions, preferably in the same experiment;the referred-to-plant can be a plant from which it was derived, e.g.,the progenitor plant, the progenitor parent, the recurrent parent, theplant used for tissue- or cell culture, etc. A physiological ormorphological characteristic can be a numerical characteristic or anon-numerical characteristic. In one aspect, a plant has “all but one,two or three of the physiological and morphological characteristics” ofa referred-to-plant, or “all the physiological and morphologicalcharacteristics” of Table 1-3 or “all or all but one, two or three ofthe physiological and morphological characteristics” of Table 1-3.

The physiological and/or morphological characteristics mentioned aboveare commonly evaluated at significance levels of 1%, 5%, or 10% if theyare numerical (quantitative), or for having an identical degree (ortype) if not numerical (not quantitative), if measured under the sameenvironmental conditions. For example, a progeny plant or a single locusconverted plant or a mutated plant of lettuce variety NUN 09161 LTL mayhave one or more (or all) of the essential physiological and/ormorphological characteristics of said variety listed in Table 1-3, asdetermined at the 5% significance level (i.e., p<0.05) for quantitativecharacteristics and determined by type or degree for non-quantitativecharacteristics, when grown under the same environmental conditions.

“Distinguishing characteristics” or “distinguishing morphological and/orphysiological characteristics” refers herein to the characteristicswhich distinguish a new variety from other lettuce varieties (i.e., aredifferent), when grown under the same environmental conditions. Thedistinguishing characteristics between lettuce variety NUN 09161 LTL andthe Reference Variety are described Table 4. When comparing lettucevariety NUN 09161 LTL with different varieties, the distinguishingcharacteristics will be different. In one aspect, the distinguishingcharacteristics may therefore include at least one, two, three or more(or all) of the characteristics listed in Table 1-3. All numericaldistinguishing characteristics are statistically significantly differentat p<0.05 between lettuce variety NUN 09161 LTL and the other variety(e.g., the Reference Variety).

Lettuce Variety NUN 09161 LTL has the following distinguishingcharacteristics when compared to the Reference Variety as shown in Table4: 1) larger plant spread of frame leaves; 2) shorter plant; 3) longerbase of head to apex (core length); 4) larger base of head (core width);5) fewer mature leaf divisions; 6) yellowish hue green color of matureleaves; 6) glossier mature leaves; 7) stronger undulation of margin ofmature leaves; and 8) medium depth of incisions of margin of matureleaves, when determined at the 5% significance level for numericalcharacteristics and determined by type or degree for non-numericalcharacteristics when grown under the same environmental conditions.

Thus, a lettuce plant “comprising the distinguishing characteristics oflettuce variety NUN 09161 LTL” (such as a progeny plant) refers hereinto a plant which does not differ from said variety in the distinguishingcharacteristics above. Therefore, in one aspect, the disclosure providesa plant that does not differ from lettuce variety NUN 09161 LTL in thedistinguishing characteristics above.

Similarity and differences between two different plant lines orvarieties can be determined by comparing the number of morphologicaland/or physiological characteristics that are the same (i.e.,statistically not significantly different) or that are different (i.e.,statistically significantly different) between the two plant lines orvarieties when grown under the same environmental conditions. Anumerical characteristic is considered to be “the same” when the valuefor a numeric characteristic is not significantly different at the 1%(p<0.01) or 5% (p<0.05) significance level, using T-test, a standardmethod known to the skilled person. A non-numerical characteristic isconsidered “the same” when the values have the same “degree” or “type”when scored using USDA and/or UPOV descriptors, for plants grown underthe same environmental conditions.

In one aspect, a statistical analysis of the quantitativecharacteristics showing the degree of significance may be provided.Statistical significance is the likelihood that a relationship betweentwo or more variables is caused by something other than chance, i.e.,that the differences in the means for quantitative characteristics oflettuce variety NUN 09161 LTL and the Reference Variety are significantor due to chance. For the purpose of proving differences or distinctionbetween lettuce variety NUN 09161 LTL and the Reference Variety, ap-value of 5% (or 0.05) or lower is considered statisticallysignificant. This means that there is only a 5% probability that theobserved result could have happened just by chance or random variation.

The statistical analysis is drawn from a small sample of at least 15plants or plant parts of lettuce variety NUN 09161 LTL and the ReferenceVariety. Statistical points or parameters such as mean, minimum, median,maximum, and standard deviation are collected from the sample data toanalyze where the average is, how varied the data set is, and whetherthe data is skewed. For the purpose of determining whether the result ofa data set is statistically significant, a T-test is used, a statisticaltool for proving significance in the means of the two groups at 5%significance level (p-value of 5% or 0.05).

The term “traditional breeding techniques” encompasses herein crossing,selfing, selection, doubled haploid production, embryo rescue,protoplast fusion, marker assisted selection, mutation breeding etc. asknown to the breeder (e.g., methods other than geneticmodification/transformation/transgenic methods, e.g., gene editing), bywhich, for example, a genetically heritable trait can be transferredfrom one lettuce line or variety to another.

“Variety” or “cultivar” means a plant grouping within a single botanicaltaxon of the lowest known rank.

“Plant line” is, for example, an inbred variety or a breeding line whichcan be used to develop one or more varieties. Both are typically highlyhomozygous. Progeny obtained by selfing such a plant line has the samephenotype as its parents.

“Inbred variety” refers to an inbred (nearly homozygous) line or seedsthereof. For example, the (nearly homozygous) plant is self-pollinatedor the (nearly homozygous) female parent is pollinated with pollen ofthe same plant line to produce inbred seeds on the female parent.

“Progeny” as used herein refers to a plant obtained from a plantdesignated NUN 09161 LTL. A progeny may be obtained by regeneration ofcell culture or tissue culture or parts of a plant of said variety orselfing of a plant of said variety or by producing seeds of a plant ofsaid variety. In further aspects, progeny may also encompass plantsobtained from crossing of at least one plant of said variety withanother lettuce plant of the same or another variety or line, or wildlettuce plants. A progeny may comprise a mutation or a transgene. A“first generation progeny” is the progeny is directly derived from,obtained from, obtainable from or derivable from the parent plant by,e.g., traditional breeding methods (selfing and/or cross-pollinating) orregeneration (optionally combined with transformation or mutation).Thus, a plant of lettuce variety NUN 09161 LTL is the male parent, thefemale parent, or both of a first generation progeny of lettuce varietyNUN 09161 LTL. Progeny may have all of the physiological andmorphological characteristics of lettuce variety NUN 09161 LTL whengrown under the same environmental conditions. Using methods such asbackcrossing, recurrent selection, mutation or transformation, one ormore specific characteristics may be introduced into said variety, or toa plant comprising all but one, two, or three of the morphological andphysiological characteristics of lettuce variety NUN 09161 LTL.

“Tissue Culture” refers to a composition comprising isolated cells ofthe same or a different type or a collection of such cells organizedinto parts of a plant. Tissue culture of various tissues of lettuce andregeneration of plants therefrom is well known and widely published(see, e.g., Teng et al., HortScience. 1992, 27(9): 1030-1032; Teng etal., HortScience. 1993, 28(6): 669-1671; Zhang et al., Journal ofGenetics and Breeding. 1992, 46(3): 287-290). Similarly, methods ofpreparing cell cultures are known in the art.

“Vegetative propagation,” “vegetative reproduction,” or “clonalpropagation” are used interchangeably herein and mean the method oftaking part of a plant and allowing that plant part to form at leastroots, and also refer to the plant or plantlet obtained by that method.Optionally, the vegetative propagation is grown into a mature plant.

“Regeneration” refers to the development of a plant from cell culture ortissue culture or vegetative propagation.

“Crossing” refers to the mating of two parent plants. The termencompasses “cross-pollination” and “selfing.”

“Selfing” refers to self-pollination of a plant, i.e., the transfer ofpollen from the anther to the stigma of the same plant.

“Cross-pollination” refers to the fertilization by the union of twogametes from different plants.

“Backcrossing” is a traditional breeding technique used to introduce atrait into a plant line or variety. The plant containing the trait iscalled the donor plant and the plant into which the trait is transferredis called the recurrent parent. An initial cross is made between thedonor parent and the recurrent parent to produce a progeny plant.Progeny plants which have the trait are then crossed to the recurrentparent. After several generations of backcrossing and/or selfing therecurrent parent comprises the trait of the donor. The plant generatedin this way may be referred to as a “single trait converted plant.” Thetechnique can also be used on a parental line of a hybrid.

The terms “gene converted,” “conversion plant,” or “single locusconverted plant” in this context refer to lettuce plants which aredeveloped by traditional backcrossing techniques, e.g., backcrossing, orvia genetic engineering (e.g., gene editing) or through mutationbreeding, wherein essentially all of the desired morphological andphysiological characteristics of parent variety or line are recovered,in addition to the one or more characteristics introduced into theparent via e.g., backcrossing technique (optionally including reversesynthesis of breeding line). It is understood that only the addition ofa further characteristic (e.g., addition of gene conferring a furthercharacteristic, such as a disease resistance gene), but also thereplacement/modification of an existing characteristic by a differentcharacteristic is encompassed herein (e.g., a mutant allele of a genecan modify the phenotype of a characteristic).

Likewise, a “Single Locus Converted (Conversion) Plant” refers to plantsdeveloped by plant breeding techniques comprising or consisting ofmutation breeding and/or by genetic transformation (e.g., gene editing)and/or by traditional breeding techniques, such as backcrossing, whereinessentially all of the desired morphological and physiologicalcharacteristics of a lettuce variety are recovered in addition to thecharacteristics of the single locus having been transferred into thevariety via the above-mentioned techniques, or wherein a morphologicaland physiological characteristic of the variety has beenreplaced/modified in the variety. In case of a hybrid, the gene may beintroduced, or modified, in the male or female parental line.

“Transgene” refers to a genetic locus comprising a DNA sequence whichhas been introduced into the genome of a lettuce plant bytransformation. A plant comprising a transgene stably integrated intoits genome is referred to as “transgenic plant.”

“Locus” (plural loci) refers to the specific location of a gene or DNAsequence on a chromosome. A locus may confer a specific trait.

“Genotype” refers to the genetic composition of a cell or organism.

“Allele” refers to one or more alternative forms of a gene locus. All ofthese loci relate to one trait. Sometimes, different alleles can resultin different observable phenotypic traits, such as differentpigmentation. However, many variations at the genetic level result inlittle or no observable variation.

As used herein, the terms “resistance” and “tolerance” are usedinterchangeably to describe plants that show no symptoms orsignificantly reduced symptoms to a specified biotic pest, pathogen,abiotic influence or environmental condition compared to a susceptibleplant. These terms are optionally also used to describe plants showingsome symptoms but that are still able to produce marketable product withan acceptable yield.

“Average” refers herein to the arithmetic mean.

The term “mean” refers to the arithmetic mean of several measurements.The mean, if not indicated otherwise within this application, refers tothe arithmetic mean of measurements on at least 15 different, randomlyselected plants of a variety or line.

DETAILED DESCRIPTION OF VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure relates to the plant of lettuce variety NUN 09161 LTL,wherein a representative sample of seeds of said variety has beendeposited under Budapest Treaty, with Accession Number NCIMB 43998. NUN09161 LTL is a multileaf green compact lettuce variety and is suitablefor growing in the open field.

In another aspect, the plant of lettuce variety NUN 09161 LTL, or partthereof, or a progeny thereof comprises resistance Downy Mildew (Bremialactucae) Isolates B1:16-37EU and Nasonovia ribisnigri Biotype Nr:0,measured according to UPOV standards described in TG/13/11.

The disclosure also provides a lettuce plant or part thereof having allof the physiological and morphological characteristics of lettucevariety NUN 09161 LTL, when grown under the same environmentalconditions.

The disclosure also provides a plant of lettuce variety NUN 09161 LTL ora progeny plant thereof comprises all of the morphological and/orphysiological characteristics (i.e., average values, as indicated on theUSDA Objective Description of variety—lettuce (unless indicatedotherwise)) as shown in Table 1-3, when the characteristics aredetermined at the 5% significance level for plants grown under the sameenvironmental conditions. A part of this plant is also provided.

The disclosure further provides a lettuce plant which does not differfrom the physiological and morphological characteristics of the plant oflettuce variety NUN 09161 LTL, as determined at the 1%, 2%, 3%, 4%, or5% significance level, when grown under the same environmentalconditions. In a particular aspect, the plants are measured in the sametrial (e.g., the trial is conducted as recommended by the USDA or UPOV).The disclosure also comprises part of said plant, preferably a leaf or apart thereof.

The morphological and/or physiological differences between two differentindividual plants described herein (e.g., between lettuce variety NUN09161 LTL and a progeny of lettuce variety NUN 09161 LTL) or between aplant of lettuce variety NUN 09161 LTL or progeny of said variety, or aplant having all, or all but 1, 2, or 3 of the physiological andmorphological characteristics of lettuce variety NUN 09161 LTL (or all,or all but 1, 2, or 3 of the characteristics as listed in Table 1-3) andanother known variety can easily be established by growing said varietynext to the each other or next to the other variety (e.g., in the samefield, under the same environmental conditions), preferably in severallocations which are suitable for said lettuce cultivation, and measuringmorphological and/or physiological characteristics of a number of plants(e.g., to calculate an average value and to determine the variationrange/uniformity within the variety). For example, trials can be carriedout in Acampo Calif., USA (N 38 degrees 07′261″/W 121 degrees 18′ 807″,USA), whereby various characteristics, for example, maturity, leafshape, size and texture, leaf color and glossiness, bolt shape, surfaceand length, flower size and color, head weight, disease resistance,insect resistance, and resistance to physiological stress can bemeasured and directly compared for species of lettuce.

Thus, the disclosure comprises a lettuce plant having one, two, or threeof the physiological and/or morphological characteristics which aredifferent from those of the plant of lettuce variety NUN 09161 LTL, andwhich otherwise has all of the physiological and morphologicalcharacteristics of the plant of lettuce variety NUN 09161 LTL, whendetermined (e.g., at the 5% significance level for plants forquantitative characteristics or determined by type for non-quantitativecharacteristics) grown under the same environmental conditions. In oneaspect, the different characteristic(s) is/are a result of breeding withlettuce variety NUN 09161 LTL and selection of a progeny plantcomprising one, two, or three characteristics which are different thanin lettuce variety NUN 09161 LTL. In another aspect, the differentcharacteristic is the result of a mutation (e.g., spontaneous mutationor a human induced mutation through e.g., targeted mutagenesis ortraditional mutagenesis such as chemically or radiation inducedmutagenesis), or it is the result of transformation.

The disclosure further relates to lettuce variety NUN 09161 LTL, whichwhen compared to Reference Variety has the following distinguishingcharacteristics as shown in Table 4: 1) larger plant spread of frameleaves; 2) shorter plant; 3) longer base of head to apex (core length);4) larger base of head (core width); 5) fewer mature leaf divisions; 6)yellowish hue green color of mature leaves; 6) glossier mature leaves;7) stronger undulation of margin of mature leaves; and 8) medium depthof incisions of margin of mature leaves, when determined at the 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions.

The disclosure also relates to a seed of lettuce variety NUN 09161 LTL,wherein a representative sample of said seed has been deposited underthe Budapest Treaty with Accession Number NCIMB 43998.

In another aspect, a seed of hybrid variety NUN 09161 LTL is obtainableby crossing the male parent of lettuce variety NUN 09161 LTL with thefemale parent of lettuce variety NUN 09161 LTL and harvesting the seedsproduced on the female parent. The resultant seed of said variety can begrown to produce plants of said variety.

In another aspect, the disclosure provides a plant grown from a seed oflettuce variety NUN 09161 LTL and a plant part thereof.

The disclosure also provides a lettuce head and/or a lettuce leafproduced on a plant grown from a seed of lettuce variety NUN 09161 LTL.

In another aspect, the disclosure provides for a plant part of lettucevariety NUN 09161 LTL, preferably a head or a leaf, a representativesample of seed from said variety has been deposited under AccessionNumber NCIMB 43998.

Also provided is a plant of lettuce variety NUN 09161 LTL, or a head ora leaf or other plant part thereof, produced from a seed, wherein arepresentative sample of said seeds has been deposited under theBudapest Treaty, with Accession Number NCIMB 43998.

Also provided is a plant part obtained from variety NUN 09161 LTL,wherein said plant part is a leaf, a part of a leaf, a head, a part of ahead, a fruit, a part of a fruit, pollen, an ovule, a cell, a petiole, ashoot or a part thereof, a stem or a part thereof, a root or a partthereof, a root tip, a cutting, a seed, a part of a seed, seed coat, oranother maternal tissue which is part of a seed grown on a said variety,a hypocotyl, a cotyledon, a pistil, an anther, or a flower or a partthereof. Such plant parts may be suitable for sexual reproduction (e.g.,pollen, a flower, an ovary, an ovule, an embryo, etc.), vegetativereproduction (e.g., a cutting, a root, a stem a cell, a protoplast, aleaf, a cotyledon, a meristem, etc.), or tissue culture (e.g., a leaf, apollen, an embryo, a cotyledon, a hypocotyl, a cell, a root, a root tip,an anther, a flower, a seed, a stem, etc.). Heads and leaves areparticularly important plant parts.

In a further aspect, the plant part obtained from variety NUN 09161 LTLis a cell, optionally a cell in a cell or tissue culture. The cell maybe grown into a plant of lettuce variety NUN 09161 LTL. A part oflettuce variety NUN 09161 LTL (or of a progeny of that variety or of aplant having all of the physiological and morphological characteristicsbut one, two, or three of lettuce variety NUN 09161 LTL) furtherencompasses any cells, tissues, organs obtainable from the seedlings orplants in any stage of maturity.

The disclosure also provides a tissue or cell culture comprisingregenerable cells of lettuce variety NUN 09161 LTL. Such tissue culturecan, for example, be grown on plates or in liquid culture or be frozenfor long term storage. The cells of lettuce variety NUN 09161 LTL usedto start the culture can be any plant part suitable for vegetativereproduction, or, in a particular aspect, can be one or more of: anembryo, a meristem, a cotyledon, a hypocotyl, a pollen, a leaf, ananther, a root, a root tip, a petiole, a flower, a fruit, a seed, or astem. In another particular aspect, the tissue culture does not containsomaclonal variation or has reduced somaclonal variation. The skilledperson is familiar with methods to reduce or prevent somaclonalvariation, including regular reinitiation.

In another aspect, the disclosure provides a lettuce plant regeneratedfrom the tissue or cell culture of lettuce variety NUN 09161 LTL,wherein the regenerated plant is not different from lettuce variety NUN09161 LTL in all, or all but one, two or three, of the physiological andmorphological characteristics (determined at the 5% significance levelwhen grown under the same environmental conditions. Optionally, theplant has one, two, or three physiological or morphologicalcharacteristic that is different from lettuce variety NUN 09161 LTL,wherein the difference or modification is effected by mutation ortransformation with a transgene.

In another aspect, the disclosure provides a lettuce plant regeneratedfrom the tissue or cell culture of variety NUN 09161 LTL, wherein theplant has all or all but one, two, or three of the physiological andmorphological characteristics of said variety determined at the 5%significance level when grown under the same environmental conditions.Similarity or difference of a characteristic is determined by measuringthe characteristics of a representative number of plants grown under thesame environmental conditions, determining whether type/degreecharacteristics are the same and determining whether numericalcharacteristics are different at the 5% significance level.

Lettuce variety NUN 09161 LTL, or its progeny, or a plant having allphysiological and/or morphological characteristics or all but one, two,or three which are different from those of lettuce variety NUN 09161LTL, can also be reproduced using vegetative reproduction methods.Therefore, the disclosure provides for a method of producing a plant orplant part of lettuce variety NUN 09161 LTL, comprising vegetativereproduction of lettuce variety NUN 09161 LTL. Vegetative propagationcomprises regenerating a whole plant from a plant part of lettucevariety NUN 09161 LTL or from a progeny or from a plant having all ofthe physiological and morphological characteristics of said variety orall but one, two, or three different characteristics, such as a cutting,a cell culture, or a tissue culture.

The disclosure also provides methods of vegetatively propagating a partof the plant of variety NUN 09161 LTL. In certain aspects, the methodcomprises: (a) cultivating tissue or cells capable of being propagatedfrom lettuce variety NUN 09161 LTL to obtain proliferated shoots; and(b) rooting said proliferated shoots, to obtain rooted plantlets. Steps(a) and (b) may also be reversed, i.e., first cultivating said tissue toobtain roots and then cultivating the tissue to obtain shoots, therebyobtaining rooted plantlets. The rooted plantlets may then be furthergrown, to obtain plant. In one aspect, the method further comprises (c)growing plants from said rooted plantlets. Therefore, the method alsocomprises regenerating a whole plant from a part of lettuce variety NUN09161 LTL. In a particular aspect, the part to be propagated is acutting, a cell culture, or a tissue culture.

The disclosure also provides for a vegetatively propagated plant ofvariety NUN 09161 LTL (or from progeny of lettuce variety NUN 09161 LTLor from a plant having all but one, two, or three of the physiologicaland/or morphological characteristics of lettuce variety NUN 09161 LTL),wherein the plant has all of the physiological and morphologicalcharacteristics of lettuce variety NUN 09161 LTL when thecharacteristics are determined at the 5% significance level for plantsgrown under the same environmental conditions. In another aspect, thepropagated plant has all but one, two, or three of the physiological andmorphological characteristics of lettuce variety NUN 09161 LTL when thecharacteristics are determined at the 5% significance level for plantsgrown under the same environmental conditions. A part of said propagatedplant or said propagated plant with one, two, or three differences isalso provided. In another aspect, the propagated plant has all or allbut one, two, or three of the physiological and morphologicalcharacteristics of lettuce variety NUN 09161 LTL (e.g., as listed inTable 1-3).

In another aspect, the disclosure provides a method for producing aplant part, preferably a head or a leaf, comprising growing a plant oflettuce variety NUN 09161 LTL until it develops at least one leaf ordevelops a head, and optionally collecting the head or leaf. Preferably,the head or leaf is collected at harvest maturity. In another aspect,the leaf is collected at baby leaf stage.

In another aspect, a plant of lettuce variety NUN 09161 LTL can beproduced by seeding directly in the soil (e.g., field) or by germinatingthe seeds in controlled environment conditions (e.g., greenhouses,hydroponic cultures, etc.) and optionally transplanting the seedlingsinto the field (see, e.g., Gonai et al., J. of Exp. Bot., 55(394): 111,2004; Turini, et. al., University of California Agricultural and NaturalResources, 2011, Publication 7215, 1-6; Smith, et. al., University ofCalifornia Agricultural and Natural Resources, 2011, Publication 7216,1-6). Lettuce may also be grown in tunnels. Moreover, said variety canbe grown in hydroponic cultures as described in, e.g., US 2008/0222949,which is herein incorporated by reference in its entirety, and theskilled person is familiar with various types of hydroponic cultures.Alternatively, seed of lettuce variety NUN 09161 LTL may be grown onpeat block for use as root ball lettuce. Furthermore, said variety maybe combined with 1, 2 or 3 different lettuce varieties to be grown as“composite lettuce” (see, e.g., EP 1197137, which is herein incorporatedby reference in its entirety).

In another aspect, the plant and plant parts of lettuce variety NUN09161 LTL and progeny of said variety are provided, e.g., grown fromseeds, produced by sexual or vegetative reproduction, regenerated fromthe above-described plant parts, or regenerated from cell or tissueculture of the lettuce variety NUN 09161 LTL, in which the reproduced(seed propagated or vegetatively propagated) plant has all of thephysiological and morphological characteristics of lettuce variety NUN09161 LTL, e.g., as listed in Table 1-3. In one aspect, said progeny oflettuce variety NUN 09161 LTL can be modified in one, two, or threecharacteristics, in which the modification is a result of mutagenesis ortransformation with a transgene.

In other aspects, the disclosure provides for a progeny plant of varietyNUN 09161 LTL such as a progeny plant obtained by further breeding oflettuce variety NUN 09161 LTL. Further breeding with said varietyincludes selfing and/or cross-pollinating lettuce variety NUN 09161 LTLwith another lettuce plant or variety one or more times. In a particularaspect, the disclosure provides for a progeny plant that retains all ofthe morphological and physiological characteristics of lettuce varietyNUN 09161 LTL, optionally all or all but one, two, or threecharacteristics as listed in Table 1-3, determined at the 5%significance level for numerical characteristics, when grown under thesame environmental conditions. In another aspect, the progeny is a firstgeneration progeny, i.e., the ovule or the pollen (or both) used in thecrossing is an ovule or pollen of lettuce variety NUN 09161 LTL, wherethe pollen comes from an anther of lettuce variety NUN 09161 LTL and theovule comes from an ovary of lettuce variety NUN 09161 LTL.

In still another aspect, the disclosure provides a method of producing alettuce plant, comprising crossing a plant of lettuce variety NUN 09161LTL with a second lettuce plant at least once, allowing the seed todevelop and optionally harvesting said progeny seed. The skilled personcan select a progeny lettuce plant from said crossing. Optionally, theprogeny (grown from the progeny seed) is crossed twice, thrice, or four,six or seven times, and allowed to set seed. In one aspect, the first“crossing” further comprises planting seeds of a first and a secondparent lettuce plant, often in proximity so that pollination will occur;for example, mediated by insect vectors. Alternatively, pollen can betransferred manually. Where the plant is self-pollinated, pollinationmay occur without the need for direct human intervention other thanplant cultivation. After pollination, the plant can produce seed.

The disclosure also provides a method for collecting pollen of lettucevariety NUN 09161 LTL, comprising collecting pollen from a plant ofvariety NUN 09161 LTL. Alternatively, the method comprises growing aplant of lettuce variety NUN 09161 LTL until at least one flowercontains pollen and collecting the pollen. In a particular aspect, thepollen is collected when it is mature or ripe. A suitable method forcollecting pollen comprises collecting anthers or the part of the antherthat contains pollen, for example, by cutting the anther or the part ofthe anther off. Pollen can be collected in a container. Optionally,collected pollen can be used to pollinate a lettuce flower.

In yet another aspect, the disclosure provides a method of producing alettuce plant, comprising selfing a plant of variety NUN 09161 LTL oneor more times, and selecting a progeny plant from said selfing. In oneaspect, the progeny plant retains all or all but one, two, or three ofthe physiological and morphological characteristics of lettuce varietyNUN 09161 LTL when grown under the same environmental conditions. In adifferent aspect, the progeny plant comprises all (or all but one, twoor three) of the physiological and morphological characteristic oflettuce variety NUN 09161 LTL of Table 1-3.

The disclosure also provides a method for developing a lettuce plant ina lettuce breeding program, using lettuce variety NUN 09161 LTL, or itsparts as a source of plant breeding material. Suitable plant breedingtechniques are recurrent selection, backcrossing, pedigree breeding,mass selection, mutation breeding, genetic marker enhanced selection,and/or genetic transformation. In one aspect, the method comprisescrossing lettuce variety NUN 09161 LTL or progeny of said variety, or aplant comprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of lettuce variety NUN 09161 LTL (e.g., aslisted in Table 1-3) with a different lettuce plant, and wherein one ormore offspring of the crossing are subject to one or more plant breedingtechniques: recurrent selection, backcrossing, pedigree breeding, massselection, mutation breeding, genetic marker enhanced selection, genetictransformation (see, e.g., Brotman et al., Theor Appl Genet (2002)104:1055-1063). For breeding methods in general, see, e.g., Acquaah,Principles of Plant Genetics and Breeding, 2007, Blackwell Publishing,ISBN-13: 978-1-4051-3646-4.

In one aspect, pedigree selection is used as a breeding method fordeveloping a lettuce variety. Pedigree selection is also known as the“Vilmorin System of Selection,” see, e.g., Allard, John Wiley & Sons,Inc., 1999, pp. 64-67. In general, selection is first practiced among F2plants. In the next season, the most desirable F3 lines are firstidentified, then desirable F3 plants within each line are selected. Thefollowing season and in all subsequent generations of inbreeding, themost desirable families are identified first, then desirable lineswithin the selected families are chosen. A family refers to lines thatwere derived from plants selected from the same progeny from thepreceding generation.

Using pedigree method, two parents may be crossed using an emasculatedfemale and a pollen donor (male) to produce F1 offspring. Lettuce is anobligate self-pollination species, which means that pollen is shedbefore stigma emergence, assuring 100% self-fertilization. Therefore, inorder to optimize crossing, a method of misting may be used to wash thepollen off prior to fertilization to assure crossing or hybridization.The F1 may be self-pollinated to produce segregating F2 generation.Individual plants may then be selected which represent the desiredphenotype in each generation (F3, F4, F5, etc.) until the traits arehomozygous or fixed within a breeding population.

Thus, progeny in connection with pedigree selection are either thegeneration (seeds) produced from the first cross (F1) or selfing (S1),or any further generation produced by crossing and/or selfing (F2, F3,F4, F5, F6, F7, etc.) and/or backcrossing (BC1, BC2, BC3, BC4, BC5, BC6,BC7, etc.) one or more selected plants of the F1 and/or S1 and/or BC1generation (or plants of any further generation, e.g., the F2) withanother lettuce plant (an/or with wild relative of lettuce).

In yet another aspect, the disclosure provides for a method of producinga new lettuce plant. The method comprises crossing lettuce variety NUN09161 LTL, or a plant of comprising all but 1, 2, or 3 of themorphological and physiological characteristics of said lettuce variety(as listed in Table 1-3), or a progeny plant thereof, either as male oras female parent, with a second lettuce plant (or a wild relative oflettuce) one or more times, and/or selfing a lettuce plant of varietyNUN 09161 LTL, or a progeny plant thereof, one or more times, andselecting progeny from said crossing and/or selfing. The second lettuceplant may, for example, be a line or variety of the species Lactucasativa, or other Lactuca species.

In a further aspect, lettuce variety NUN 09161 LTL is used in crosseswith other, different, lettuce varieties to produce first generation(F1) lettuce hybrid seeds and plants with superior characteristics. In aparticular aspect, the disclosure provides a method a producing a hybridlettuce seed comprising crossing a first parent lettuce plant with asecond parent lettuce plant and harvesting the resultant seed, in whichthe first parent lettuce plant or second parent lettuce plant is lettucevariety NUN 09161 LTL. Also provided is a hybrid lettuce seed producedfrom crossing a first parent lettuce plant with a second parent lettuceplant and harvesting the resultant hybrid lettuce seed, wherein saidfirst parent lettuce plant or second parent lettuce plant is lettucevariety NUN 09161 LTL. In a further aspect, the hybrid lettuce plantproduce from the hybrid lettuce seed is provided.

The morphological and physiological characteristics (and thedistinguishing characteristics) of lettuce variety NUN 09161 LTL areprovided, for example, in Table 1-3. Encompassed herein is also a plantobtainable from lettuce variety NUN 09161 LTL (e.g., by selfing and/orcrossing and/or backcrossing with said variety and/or progeny of saidvariety) comprising all or all but one, two or three of thephysiological and morphological characteristics of lettuce variety NUN09161 LTL listed in Table 1-3 as determined at the 5% significance levelfor numerical characteristics or identical for non-numericalcharacteristics when grown under the same environmental conditionsand/or comprising one or more (or all; or all except one, two or three)when grown under the same environmental conditions. The morphologicaland/or physiological characteristics may vary somewhat with variation inthe environment (e.g., temperature, light intensity, day length,humidity, soil, fertilizer use), which is why a comparison under thesame environmental conditions is preferred. Colors can best be measuredagainst the Royal Horticultural Society (RHS) Chart. Also, at-harvestand/or post-harvest characteristics of heads or leaves can be compared,such as cold storage holding quality, post-harvest leaf crispness andleaf browning or pinking after cutting can be measured using knownmethods.

In another aspect, a seed of inbred variety NUN 09161 LTL is obtainableby selfing the variety and harvesting the seeds produced. The resultantseeds can be grown to produce plants of said variety.

In still another aspect, the disclosure provides a method of producing aplant derived from lettuce variety NUN 09161 LTL, the method comprising:(a) preparing a progeny plant derived from lettuce variety NUN 09161 LTLby crossing a plant of variety NUN 09161 LTL either as a male or femaleparent with a second plant or selfing lettuce variety NUN 09161 LTL orvegetative reproduction of lettuce variety NUN 09161 LTL, and (b)collecting seeds from said crossing or selfing or regenerating a wholeplant from the vegetative cell- or tissue culture. Also provided areseeds and/or plants obtained by this method. All plants produced usinglettuce variety NUN 09161 LTL as a parent are within the scope of thedisclosure, including plant parts derived from lettuce variety NUN 09161LTL.

In further aspects, the method comprises growing a progeny plant of asubsequent generation from said seed of a progeny plant of a subsequentgeneration and crossing the progeny plant of a subsequent generationwith itself or a second plant and repeating the steps for an additional3-10 generations to produce a plant derived from lettuce variety NUN09161 LTL. The plant derived from lettuce variety NUN 09161 LTL may bean inbred line and the aforementioned repeating crossing steps may bedefined as comprising sufficient inbreeding to produce the inbred line.By selecting plants having one or more desirable traits, a plant derivedfrom lettuce variety NUN 09161 LTL is obtained which has some of thedesirable traits of the line as well as potentially other selectedtraits.

The disclosure provides for methods of producing plants which retain allthe morphological and physiological characteristics of a plant describedherein. The disclosure also provides for methods of producing a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of lettuce variety NUN 09161 LTL (e.g., aslisted in Table 1-3), but which are still genetically closely related tosaid variety. The relatedness can, for example, be determined byfingerprinting techniques (e.g., making use of isozyme markers and/ormolecular markers such as Single-nucleotide polymorphism (SNP) markers,amplified fragment length polymorphism (AFLP) markers, microsatellites,minisatellites, Random Amplified Polymorphic DNA (RAPD) markers,restriction fragment length polymorphism (RFLP) markers and others). Aplant is “closely related” to lettuce variety NUN 09161 LTL if its DNAfingerprint is at least 80%, 90%, 95% or 98% identical to thefingerprint of lettuce variety NUN 09161 LTL. In a particular aspect,AFLP markers are used for DNA fingerprinting (see, e.g., Vos et al.1995, Nucleic Acid Research 23: 4407-4414). A closely related plant mayhave a Jaccard's Similarity index of at least about 0.95 or 0.96 or more(see, e.g., “Guidelines for the Handling of a Dispute on EssentialDerivation in Lettuce” atworldseed.org/wp-content/uploads/2015/10/GuidelinesEDV_Lettuce_2004.pdf). the disclosure also provides a plant and avariety obtained or selected by applying these methods on lettucevariety NUN 09161 LTL. Such a plant may be produced by crossing and/orselfing, or alternatively, a plant may simply be identified and selectedamongst plants of said variety, or progeny of said variety, e.g., byidentifying a variant within lettuce variety NUN 09161 LTL or progeny ofsaid variety (e.g., produced by selfing) which variant differs fromlettuce variety NUN 09161 LTL in one, two or three of the morphologicaland/or physiological characteristics (e.g., characteristics listed inTable 1-3). In one aspect, the disclosure provides a plant of lettucevariety NUN 09161 LTL having a Jaccard's Similarity index with saidvariety of at least 0.95, 0.96, 0.97, 0.98 or even at least 0.99.

In some aspects, the disclosure provides a lettuce plant comprisinggenomic DNA having at least 95%, 96%, 97%, 98%, or 99% sequence identitycompared to the genomic DNA sequence of a plant of lettuce variety NUN09161 LTL as deposited under Accession Number NCIMB 43998. In someaspects, the lettuce plant further comprises all or all but one, two, orthree of the physiological and morphological characteristics of lettucevariety NUN 09161 LTL (e.g., as listed in Table 1-3). In other aspects,the lettuce plant comprises the distinguishing characteristics oflettuce variety NUN 09161 LTL.

For the purpose of this disclosure, the “sequence identity” ornucleotide sequences, expressed as a percentage, refers to the number ofpositions in the two optimally aligned sequences which have identicalresidues (×100) divided by the number of positions compared. A gap,i.e., a position in the pairwise length where a residue is present inone sequence but not in the other, is regarded as a position withnon-identical residues. A pairwise global sequence alignment of twonucleotide sequences is found by aligning the two consequences over theentire length according to the Needleman and Wunsch global alignmentalgorithm described in Needleman and Wunsch, 1970, J. Mol. Bio.48(3):443-53. A full implementation of the Needleman-Wunsch globalalignment algorithm is found in the needle program in The EuropeanMolecular Biology Open Software (see, EMBOSS, Rice, et. al., Trends inGenetics, June 2000, 16(6):276-77).

In one aspect, a plant of lettuce variety NUN 09161 LTL may also bemutated (by e.g., irradiation, chemical mutagenesis, heat treatment,etc.) and mutated seeds or plants may be selected in order to change oneor more characteristics of said variety. Methods such as TILLING(Targeting Induced Local Lesions in Genomes) may be applied to lettucepopulations in order to identify mutants.

Similarly, lettuce variety NUN 09161 LTL may be transformed andregenerated, whereby one or more chimeric genes are introduced into thevariety or into a plant comprising all but one, two, or three of thephysiological and morphological characteristics (e.g., as listed inTable 1-3). Many useful traits can be introduced into lettuce varietyNUN 09161 LTL by e.g., crossing lettuce variety NUN 09161 LTL with atransgenic lettuce plant comprising a desired transgene, as well as bydirectly introducing a transgene into lettuce variety NUN 09161 LTL bygenetic transformation techniques.

Any pest or disease resistance genes may be introduced into lettucevariety NUN 09161 LTL, progeny of said variety, or into a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of lettuce variety NUN 09161 LTL (e.g., aslisted in Table 1-3). Resistance to one or more of the followingdiseases or pests may be introduced into the plant described herein:Rhizomonas suberifaciens (Corky root rot), Bremia lactucae (Downymildew), Erysiphe cichoracearum f. sp. lactucae (Powdery mildew),Sclerotinia minor and Sclerotinia sclerotiorum (Lettuce Drop),Pseudomonas spp. (Bacterial Soft Rot), Botrytis cinerea (Grey Mold),Verticillium dahlia (Verticillium Wilt), Xanthomonas spp. (BacterialLeaf Spot), Microdochium panattonianum (Anthracnose), Fusarium oxysporumf sp. lactucae, Rhizoctonia solani (Bottom Rot), Cabbage Loopers,Lettuce Root Aphid, Myzus persicae (Green Peach Aphid), Liriomyza langei(Pea Leafminer), Liriomyza trifolii (Serpentine Leafminer), Liriomyzasativae (Vegetable Leafminer), Foxglove Aphid, Potato Aphid, BeetArmyworm, Bemisia argentifolii (Silver Whitefly), and/or Aster Yellows.Other resistance genes, against pathogenic viruses (e.g., MirafioriLettuce Big Vein Virus (LMBVV), Lettuce Infectious Yellows Virus (LIYV),Lettuce Mosaic Virus (LMV), Lettuce Necrotic Stunt Virus (LNSV),Cucumber Mosaic Virus (CMV), Tomato Bushy Stunt Virus (Dieback), TomatoSpotted Wilt Virus (TSWV), Turnip Mosaic Virus, Beet Western YellowsVirus (BWYV), Alfalfa mosaic virus (AMV)), fungi, bacteria, nematodes,insects or other pests may also be introduced. In one aspect, resistanceagainst Nasonovia ribisnigri biotype Nr:0 and/or Nr:1 may be introducedinto the plant disclosed herein. Also, any resistances to physiologicalstresses may be introduced into the plant described herein, or progenythereof or into a plant comprising all but 1, 2, or 3 or more of themorphological and physiological characteristics of said plant (e.g., aslisted in Table 1-3). Resistance against one or more of the followingmay be introduced into the plant described herein: Tip burn, Heat,Drought, Cold, Salt and/or Brown rob (Rib Discoloration/Rib Blight).

Genetic transformation may, therefore, be used to insert a selectedtransgene into the lettuce plants of the disclosure described herein ormay, alternatively, be used for the preparation of transgenic lettuceplants which can be used as a source of the transgene(s), which can beintroduced into lettuce variety NUN 09161 LTL by e.g., backcrossing. Agenetic trait which has been engineered into the genome of a particularlettuce plant may then be moved into the genome of another lettuce plant(e.g., another variety) using traditional breeding techniques which arewell known in the art. For example, backcrossing is commonly used tomove a transgene from a transformed lettuce variety into an alreadydeveloped lettuce variety and the resulting backcross conversion plantwill then comprise the transgene(s).

Any DNA sequences, whether from a different species or from the samespecies, which are inserted into the genome using transformation, arereferred to herein collectively as “transgenes.” A “transgene” alsoencompasses antisense, or sense and antisense sequences capable of genesilencing. Thus, the disclosure also relates to transgenic plants oflettuce variety NUN 09161 LTL. In some aspects, a transgenic plant oflettuce variety NUN 09161 LTL may contain at least one transgene butcould also contain at least 1, 2, 3, 4, or more transgenes.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to aregulatory element active in plant cells (e.g., promoter). Theexpression vector may contain one or more such operably linkedgene/regulatory element combinations. The vector may be in the form of aplasmid and can be used alone or in combination with other plasmids toprovide transformed lettuce plants using transformation methods toincorporate transgenes into the genetic material of the lettuceplant(s). Transformation can be carried out using standard methods, suchas Agrobacterium tumefaciens mediated transformation or biolistic,followed by selection of the transformed cells and regeneration intoplants.

Plants can also be genetically engineered, modified, or manipulated toexpress various phenotypes of horticultural interest. Through thetransformation of lettuce, the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide resistance,stress tolerance, horticultural quality, and other traits.Transformation can also be used to insert DNA sequences which control orhelp control male sterility or fertility restoration. DNA sequencesnative to lettuce as well as non-native DNA sequences can be transformedinto lettuce and used to alter levels of native or non-native proteins.Various promoters, targeting sequences, enhancing sequences, and otherDNA sequences can be inserted into the genome for the purpose ofaltering the expression of proteins. Reduction of the activity ofspecific genes (also known as gene silencing, or gene suppression) isdesirable for several aspects of genetic engineering in plants.

Genome editing is another method recently developed to geneticallyengineer plants. Specific modification of chromosomal loci or targetedmutation can be done through sequence-specific nucleases (SSNs) byintroducing a targeted DNA double strand break in the locus to bealtered. Examples of SSNs that have been applied to plants are: fingernucleases (ZFNs), transcription activator-like effector nucleases(TALENs), engineered homing endonucleases or meganucleases, andclustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9), see, e.g., Songstad, et.al., Critical Reviews in Plant Sciences, 2017, 36:1, 1-23.

Thus, the disclosure provides a method of producing a plant of lettucevariety NUN 09161 LTL having a desired trait, comprising mutating aplant or plant part of variety NUN 09161 LTL, optionally with a targetgene, and selecting a plant the desired trait, wherein the mutated plantretains all or all but one of the physiological and morphologicalcharacteristics of said variety, optionally as described Table 1-3, andcontains the desired trait, and wherein a representative sample of seedof variety NUN 09161 LTL has been deposited under Accession Number NCIMB43998. In a further aspect, the desired trait is yield, nutritionalvalue, taste, color, crunchiness, male sterility, herbicide tolerance,insect resistance, pest resistance, disease resistance, environmentalstress tolerance, modified carbohydrate metabolism, or modified proteinmetabolism, or the mutation occurs in any of the following genes:Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL, PPO.

The disclosure also provides a method for inducing a mutation in lettucevariety NUN 09161 LTL comprising:

-   -   a. exposing the seed, plant, plant part, or cell of lettuce        variety NUN 09161 LTL to a mutagenic compound or to radiation,        wherein a representative sample of seed of said variety has been        deposited under Accession Number NCIMB 43998;    -   b. selecting the seed, plant, or plant part or cell of lettuce        variety NUN 09161 LTL having a mutation; and    -   c. optionally growing and/or multiplying the seed, plant, plant        part, or cell of lettuce variety NUN 09161 LTL having the        mutation.

The disclosure also provides a method of producing a lettuce planthaving a desired trait, wherein the method comprises transforming thelettuce plant with a transgene that confers the desired trait, whereinthe transformed plant otherwise retains all of the physiological andmorphological characteristics of the plant of variety NUN 09161 LTL andcontains the desired trait. Thus, a transgenic lettuce plant is providedwhich is produced by the method described above, wherein the plantcomprises the desired trait and all of the physiological andmorphological characteristics of the plant of variety NUN 09161 LTL.

In another aspect, the disclosure provides a method of producing aprogeny of plant of variety NUN 09161 LTL further comprising a desiredtrait, said method comprising transforming the plant of lettuce varietyNUN 09161 LTL with at least one transgene that confers the desired traitand/or crossing the plant of lettuce variety NUN 09161 LTL with atransgenic lettuce plant comprising a desired transgene so that thegenetic material of the progeny that resulted from the cross containsthe desired transgene(s). Also encompassed is the progeny produced bythis method.

A desired trait (e.g., gene(s) conferring pest or disease resistance,herbicide, fungicide or insecticide tolerance, etc.) can be introducedinto lettuce variety NUN 09161 LTL, or progeny of said variety, bytransforming said variety or progeny of said variety with a transgenethat confers the desired trait, wherein the transformed plant retainsall or all but one, two or three of the phenotypic and/or morphologicaland/or physiological characteristics of lettuce variety NUN 09161 LTL orthe progeny of said variety and contains the desired trait, wherein thedesired trait is yield, nutritional value, taste, color, crunchiness,male sterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, environmental stress tolerance, modifiedcarbohydrate metabolism, or modified protein metabolism or the mutationoccurs in any of the following genes: Ferulate-5-hydrxylase, dmr1, dmr6,NCED4, PAL, PPO. In a particular aspect, the specific transgene may beany known in the art or listed herein, including, a polynucleotidesequence conferring resistance to imidazolinone, sulfonylurea,glyphosate, glufosinate, triazine, benzonitrile, cyclohexanedione,phenoxy proprionic acid and L-phosphinothricin or a polynucleotideconferring resistance to Rhizomonas suberifaciens (Corky root rot),Bremia lactucae (Downy mildew), Erysiphe cichoracearum f sp. lactucae(Powdery mildew), Sclerotinia minor and Sclerotinia sclerotiorum(Lettuce Drop), Pseudomonas spp. (Bacterial Soft Rot), Botrytis cinerea(Grey Mold), Verticillium dahlia (Verticillium Wilt), Xanthomonas spp.(Bacterial Leaf Spot), Microdochium panattonianum (Anthracnose),Fusarium oxysporum f sp. lactucae, Rhizoctonia solani (Bottom Rot),Cabbage Loopers, Lettuce Root Aphid, Myzus persicae (Green Peach Aphid),Liriomyza langei (Pea Leafminer), Liriomyza trifolii (SerpentineLeafminer), Liriomyza sativae (Vegetable Leafminer), Foxglove Aphid,Potato Aphid, Beet Armyworm, Bemisia argentifolii (Silver Whitefly),and/or Aster Yellows. Other resistance genes, against pathogenic viruses(e.g., Mirafiori Lettuce Big Vein Virus (LMBVV), Lettuce InfectiousYellows Virus (LIYV), Lettuce Mosaic Virus (LMV), Lettuce Necrotic StuntVirus (LNSV), Cucumber Mosaic Virus (CMV), Tomato Bushy Stunt Virus(Dieback), Tomato Spotted Wilt Virus (TSWV), Turnip Mosaic Virus, BeetWestern Yellows Virus (BWYV), Alfalfa mosaic virus (AMV)), fungi,bacteria, nematodes, insects or other pests may also be introduced.

By crossing and/or selfing also (one or more), single traits may beintroduced into, or modified in, lettuce variety NUN 09161 LTL (e.g.,using backcrossing breeding schemes), while retaining the remainingmorphological and physiological characteristics of said variety and/orwhile retaining one or more or all distinguishing characteristics. Asingle trait converted plant may thereby be produced. For example,disease resistance genes may be introduced, genes responsible for one ormore quality traits, yield, etc. Both single genes (e.g., dominant orrecessive) and one or more QTLs (quantitative trait loci) may betransferred into lettuce variety NUN 09161 LTL by breeding with saidvariety.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, a single trait conversion, or a desired traitinto lettuce variety NUN 09161 LTL, comprising introducing a singlelocus conversion, a single trait conversion, or a desired trait in atleast one of the parents of lettuce variety NUN 09161 LTL, and crossingthe converted parent with the other parent of lettuce variety NUN 09161LTL, to obtain seed of said variety.

In another aspect, the step of introducing a single locus conversion, asingle trait conversion, or a desired trait in at least one of theparent plants comprises:

-   -   a. crossing the parental line of lettuce variety NUN 09161 LTL,        with a second lettuce plant comprising the single locus        conversion, the single trait conversion, or the desired trait;    -   b. selecting F1 progeny plants that contain the single locus        conversion, the single trait conversion, or the desired trait.    -   c. crossing said selected progeny plants of step b) with the        parental line of step a) to produce a backcross progeny plant;    -   d. selecting backcross progeny plants comprising the single        locus conversion, the single trait conversion or the desired        trait and otherwise all or all but one, two, or three of the        physiological and morphological characteristics of the parental        line of step a) to produce selected backcross progeny plants;        and    -   e. optionally repeating steps c) and d) one or more times in        succession to produce selected second, third, or fourth, or        higher backcross progeny plants comprising the single locus        conversion, the single trait conversion, or the desired trait        and otherwise all or all but one, two, or three of the        physiological and morphological characteristics of the parental        line of step a) to produce selected backcross progeny plants,        when grown in the same environmental conditions.        The disclosure further relates to plants obtained by this        method.

Alternatively, a single trait converted plant or single locus convertedplant may be produced by:

-   -   a. obtaining a cell or tissue culture of cells of the parental        line of lettuce variety NUN 09161 LTL;    -   b. genetically transforming or mutating said cells;    -   c. growing the cells into a plant; and    -   d. optionally selecting a plant that contains the desired single        locus conversion, single trait conversion, or the desired trait.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, a single trait conversion, or a desired traitinto lettuce variety NUN 09161 LTL, comprising:

-   -   a. obtaining a combination of parental lines of lettuce variety        NUN 09161 LTL, optionally through reverse synthesis of breeding        lines;    -   b. introducing a single locus conversion, a single trait        conversion, or a desired trait in at least one of the parents of        step a); and    -   c. crossing the converted parent with the other parent of        step a) to obtain seed of lettuce variety NUN 09161 LTL.

In any of the above methods, wherein the single locus conversionconcerns a trait, the trait may be yield or pest resistance, or diseaseresistance. In one aspect, the trait is disease resistance and theresistance is conferred to Rhizomonas suberifaciens (Corky root rot),Bremia lactucae (Downy mildew), Erysiphe cichoracearum f. sp. lactucae(Powdery mildew), Sclerotinia minor and Sclerotinia sclerotiorum(Lettuce Drop), Pseudomonas spp. (Bacterial Soft Rot), Botrytis cinerea(Grey Mold), Verticillium dahlia (Verticillium Wilt), Xanthomonas spp.(Bacterial Leaf Spot), Microdochium panattonianum (Anthracnose),Fusarium oxysporum f sp. lactucae, Rhizoctonia solani (Bottom Rot),Cabbage Loopers, Lettuce Root Aphid, Myzus persicae (Green Peach Aphid),Liriomyza langei (Pea Leafminer), Liriomyza trifolii (SerpentineLeafminer), Liriomyza sativae (Vegetable Leafminer), Foxglove Aphid,Potato Aphid, Beet Armyworm, Bemisia argentifolii (Silver Whitefly),and/or Aster Yellows. Other resistance genes, against pathogenic viruses(e.g., Mirafiori Lettuce Big Vein Virus (LMBVV), Lettuce InfectiousYellows Virus (LIYV), Lettuce Mosaic Virus (LMV), Lettuce Necrotic StuntVirus (LNSV), Cucumber Mosaic Virus (CMV), Tomato Bushy Stunt Virus(Dieback), Tomato Spotted Wilt Virus (TSWV), Turnip Mosaic Virus, BeetWestern Yellows Virus (BWYV), Alfalfa mosaic virus (AMV)), fungi,bacteria, nematodes, insects or other pests may also be introduced. Inone aspect, resistance against Nasonovia ribisnigri biotype Nr:0 and/orNr:1 may be introduced into the plant disclosed herein. Also, anyresistances to physiological stresses may be introduced into the plantdescribed herein, or progeny thereof or into a plant comprising all but1, 2, or 3 or more of the morphological and physiologicalcharacteristics of said plant (e.g., as listed in Table 1-3). Resistanceagainst one or more of the following may be introduced into the plantdescribed herein: Tip burn, Heat, Drought, Cold, Salt and/or Brown rob(Rib Discoloration/Rib Blight).

The disclosure also provides a plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of lettuce variety NUN 09161 LTL, and which otherwise has allof the physiological and morphological characteristics of said variety,wherein a representative sample of seed of said variety has beendeposited under Accession Number NCIMB 43998. In particular, variantswhich differ from lettuce variety NUN 09161 LTL in none, one, two, orthree of the characteristics mentioned in Table 1-3 are encompassed.

The disclosure also provides a lettuce plant comprising at least a setof first set of the chromosomes of lettuce variety NUN 09161 LTL, asample of seed of said variety has been deposited under Accession NumberNCIMB 43998; optionally further comprising a single locus conversion ormutation, wherein said plant has essentially all of the morphologicaland physiological characteristics of the plant comprising at least afirst set of the chromosomes of said variety. In another aspect, thesingle locus conversion or mutation confers a trait, wherein the traitis yield, color, size, taste, crunchiness, enhance nutritional quality,post-harvest quality, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, or modified proteinmetabolism.

In one aspect, the disclosure provides for a haploid plant and/or adoubled haploid plant of variety NUN 09161 LTL, or a plant having allbut one, two or three physiological and/or morphological characteristicsof lettuce variety NUN 09161 LTL, or progeny of said variety. Haploidand doubled haploid (DH) plants can, for example, be produced by cell ortissue culture and chromosome doubling agents and regeneration into awhole plant. DH production chromosome doubling may be induced usingknown methods, such as colchicine treatment or the like. In one aspect,the method comprises inducing a cell or tissue culture with a chromosomedoubling agent and regenerating the cells or tissues into a whole plant.

In another aspect, the disclosure comprises a method for producingdoubled haploid cells of lettuce variety NUN 09161 LTL, comprisingmaking doubles haploid cells from haploids cells from the plant or plantpart of lettuce variety NUN 09161 LTL with a chromosome doubling agent,such as colchicine treatment (see, e.g., Nikolova andNiemirowicz-Szczytt (1996) Acta Soc Bot Pol 65:311-317).

In another aspect, the disclosure provides for haploid plants and/ordoubled haploid plants derived from lettuce variety NUN 09161 LTL that,when combined, make a set of parents of lettuce variety NUN 09161 LTL.The haploid plant and/or the doubled haploid plant of variety NUN 09161LTL can be used in a method for generating parental lines of lettucevariety NUN 09161 LTL.

The disclosure also provides methods for determining the identity ofparental lines of the plant described herein, in particular the identityof the female line. US 2015/0126380, which is hereby incorporated byreference, relates to a non-destructive method for analyzing maternalDNA of a seed. In this method, the DNA is dislodged from the seed coatsurface and can be used to collect information on the genome of thematernal parent of the seed. This method for analyzing maternal DNA of aseed, comprises the steps of contacting a seed with a fluid to dislodgeDNA from the seed coat surface, and analyzing the DNA thus dislodgedfrom the seed coat surface using methods known in the art. The skilledperson is thus able to determine whether a seed has grown on a plant ofa plant of lettuce variety NUN 09161 LTL is a progeny of said variety,because the seed coat of the seed is a maternal tissue geneticallyidentical to lettuce variety NUN 09161 LTL. Since lettuce variety NUN09161 LTL is an inbred variety, with a very high degree of homozygosity,any F1 progeny will inherit the same, predictable, set of chromosomesfrom its parent. Thus, the skilled person will also be able to identifymaternal tissues of a seed grown on an F1 progeny of lettuce variety NUN09161 LTL, using the methods described in US 2015/0126380. In anotherparticular aspect, the skilled person can determine the identity of thefemale parental line of lettuce variety NUN 09161 LTL by analyzing theseed coat of a seed of that variety. In another aspect, the skilledperson can determine whether a seed is grown on lettuce variety NUN09161 LTL.

Using methods known in the art such as “reverse synthesis of breedinglines” or “reverse breeding”, it is possible to produce parental linesfor a hybrid plant such as lettuce variety NUN 09161 LTL. A skilledperson can take any individual heterozygous plant (called a“phenotypically superior plant” in Example 2 of US 2015/0245570 herebyincorporated by reference in its entirety; lettuce variety NUN 09161 LTLis such plant) and generate a combination of parental lines (reversebreeding parental lines) that, when crossed, produce the variety NUN09161 LTL. It is not necessary that the reverse breeding parental linesare identical to the original parental lines. Such new breeding methodsare based on the segregation of individual alleles in the sporesproduced by a desired plant and/or in the progeny derived from theself-pollination of that desired plant, and on the subsequentidentification of suitable progeny plants in one generation, or in alimited number of inbred cycles. Such a method is known from US2015/0245570 or from Wijnker et al., Nature Protocols Volume: 9, Pages:761-772 (2014) DOI: doi:10.1038/nprot.2014.049. Thus, the disclosureprovides a method for producing parental lines for a hybrid organism(e.g., lettuce variety NUN 09161 LTL), comprising in one aspect: a)defining a set of genetic markers present in a heterozygous form (H) ina partially heterozygous starting organism; b) producing doubled haploidlines from spores of the starting organism; c) geneticallycharacterizing the doubled haploid lines thus obtained for the said setof genetic markers to determine whether they are present in a firsthomozygous form (A) or in a second homozygous form (B); and d) selectingat least one pair of doubled haploid lines that have complementaryalleles for at least a subset of the genetic markers, wherein eachmember of the pair is suitable as a parental line for the hybridorganism.

In another aspect, the method for producing parental lines for hybridorganisms, e.g., of lettuce variety NUN 09161 LTL, which when crossedreconstitute the genome of lettuce variety NUN 09161 LTL, comprising:

-   -   a. defining a set genetic a) markers that are present a        heterozygous form (H) in a partially heterozygous starting        organism;    -   b. producing at least one further generation from the starting        organism by self-pollination (e.g., F2 or F3 generation);    -   c. selecting at least one pair of progeny organisms in which at        least one genetic marker from the set is present in a        complementary homozygous forms (B vs. A, or A vs. B); and        optionally repeating steps b) and c) until at least one pair of        progeny organisms that have complementary alleles for at least a        subset of the genetic markers has been selected as parental        lines for a hybrid.

The disclosure also relates to a method of producing a combination ofparental lines of a plant of variety NUN 09161 LTL, comprising makingdouble haploid cells from haploid cells form said plant or a seed ofthat plant; and optionally crossing these parental lines to produce andcollect seeds. In another aspect, the disclosure relates to acombination of parental lines produced by this method. In still anotheraspect, the combination of parental lines can be used to produce a seedor plant of variety NUN 09161 LTL, when these parental lines arecrossed. In still another aspect, the disclosure relates to acombination of a parental lines from which a seed or plant having allphysiological and/or morphological characteristics of lettuce varietyNUN 09161 LTL (when the characteristics are determined at the 5%significance level for plants grown under the same environmentalconditions).

The disclosure also provides a combination of parental lines, which whencrossed, produce a seed or plant having all of the physiological and/ormorphological characteristics of lettuce variety NUN 09161 LTL, but one,two, or three which are different (when grown under the sameenvironmental conditions).

In another aspect, a combination of a male and a female parental line oflettuce variety NUN 09161 LTL can be generated by methods describedherein, for example, through reverse synthesis of breeding lines.

In another aspect, the disclosure provides a method of determining thegenotype of a plant of the invention comprising the step of detecting inthe genome (e.g., a sample of nucleic acids) of the plant at least afirst polymorphism or an allele. The skilled person is familiar withmany suitable methods of genotyping, detecting a polymorphism ordetecting an allele including SNP (Single Nucleotide Polymorphism)genotyping, restriction fragment length polymorphism identification(RFLP) of genomic DNA, random amplified polymorphic detection (RAPD) ofgenomic DNA, amplified fragment length polymorphism detection (AFLPD),polymerase chain reaction (PCR), DNA sequencing, allele specificoligonucleotide (ASO) probes, and hybridization to DNA microarrays orbeads. Alternatively, the entire genome could be sequenced. The methodmay, in certain aspects, comprise detecting a plurality of polymorphismsin the genome of the plant, for example, by obtaining a sample ofnucleic acid from a plant and detecting in said nucleic acids aplurality of polymorphisms. The method may further comprise storing theresults of the step of detecting the plurality of polymorphisms on acomputer readable medium.

Also provided is a plant part obtainable from variety NUN 09161 LTL (orfrom progeny of said variety or from a plant having all or all but one,two or three physiological and/or morphological characteristics whichare different from those of lettuce variety NUN 09161 LTL) or from avegetatively propagated plant of variety NUN 09161 LTL (or from itsprogeny or from a plant having all but one, two or three physiologicaland/or morphological characteristics which are different from those oflettuce variety NUN 09161 LTL), wherein the plant part is a leaf, aharvested leaf, a part of a leaf, a head, a harvested head, a part of ahead, pollen, an ovule, a cell, a petiole, a shoot or a part thereof, astem or a part thereof, a root or a part thereof, a root tip, a cutting,a seed, a part of a seed, seed coat or another maternal tissue which ispart of a seed grown on lettuce variety NUN 09161 LTL, or a hypocotyl, acotyledon, a pistil, an anther, or a flower or a part thereof.

A part of lettuce variety NUN 09161 LTL (or of progeny of said varietyor of a plant having all physiological and/or morphologicalcharacteristics but one, two or three which are different from those ofsaid variety) encompasses any cells, tissues, organs obtainable from theseedlings or plants, such as but not limited to: a lettuce head or apart thereof, a leaf or a part thereof, a cutting, hypocotyl, cotyledon,seed coat, pollen and the like. Such parts can be stored and/orprocessed further.

The disclosure also provides for a food product, a feed product, or aprocessed product comprising or consisting of a plant part describedherein. Preferably, the plant part is a lettuce head or leaf or a partthereof and/or an extract from a leaf or another plant part describedherein comprising at least one cell of lettuce variety NUN 09161 LTL.The food or feed product may be fresh or processed, e.g., dried,grinded, powdered, pickled, chopped, cooked, roasted, in a sauce, in asandwich, pasted, pureed or concentrated, juiced, pickled, canned,steamed, boiled, fried, blanched and/or frozen, etc.

Such plant part of lettuce variety NUN 09161 LTL can be stored and/orprocessed further. The disclosure thus also provides for a food or feedproducts comprising one or more of such parts, such as canned, chopped,cooked, roasted, in a sauce, in a sandwich, pasted, pureed orconcentrated, juiced, frozen, dried, pickled, or powdered lettuce headsor leaves from lettuce variety NUN 09161 LTL or from progeny of saidvariety, or from a derived variety, such as a plant having all or allbut one, two, or three of the physiological and/or morphologicalcharacteristics of lettuce variety NUN 09161 LTL.

The disclosure further provides for food or feed products comprisingpart, or part of progeny of lettuce variety NUN 09161 LTL, or part of aplant having all but one, two, or three of the physiological andmorphological characteristics of lettuce variety NUN 09161 LTL,comprising one or more of such parts, optionally processed (e.g.,canned, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,pureed or concentrated, juiced, frozen, dried, pickled, or powdered).

In another aspect, the plant, plant part, or seed of lettuce variety NUN09161 LTL is inside or more containers. For example, the disclosureprovides containers such as cans, boxes, crates, bags, cartons, ModifiedAtmosphere Packaging Films (e.g., biodegradable films), etc. comprisinga plant or part of a plant (fresh and/or processed) or seed of lettucevariety NUN 09161 LTL. In a particular aspect, the container comprises aplurality of seeds of lettuce variety NUN 09161 LTL, or a plurality ofplant parts of lettuce variety NUN 09161 LTL. In one aspect, a seed or aplurality of seeds of said variety are packaged into a container of anysize or type (e.g., bags, cartons, cans, etc.). The seeds may bepelleted prior to packing (to form pills or pellets) and/or may bedisinfected, primed and/or treated with various compounds, such as seedcoating or crop protection compounds. The seed produces a plant oflettuce variety NUN 09161 LTL.

In another aspect, the disclosure provides for a lettuce head or leaf ofvariety NUN 09161 LTL, or a part of a head or leaf of said variety. Thehead or leaf can be in any stage of maturity, for example, immature ormature. In another aspect, the disclosure provides for a containercomprising or consisting of a plurality of harvested lettuce heads orleaves or parts of lettuce heads or leaves of said variety, or lettuceheads or leaves of progeny thereof, or lettuce heads or leaves of aderived variety.

Marketable lettuce heads or leaves are generally sorted by size andquality after harvest. Alternatively, the lettuce heads or leaves can besorted by leaf size, shape, texture, glossiness, or color.

All documents (e.g., patent publications) are herein incorporated byreference in their entirety, including the following cited references:

-   Naktuinbow and NARO, “Calibration Manual: DUS Test for Lettuce,”    world-wide web at naktuinbow.nl-   US Department of Agriculture, Agricultural Marketing Service,    “Objective Description of Variety-Lettuce (Lactuca sativa L.)”,    world wide web at    ams.usda.gov/services/plant-variety-protection/pvpo-c-forms, under    lettuce.-   UPOV, “Guidelines for the Conduct of Tests for Distinctness,    Uniformity and Stability”, TG/13/11 (Geneva 2006, last updated 2017    Apr. 5), world-wide web at upov.int under edocs/tgdocs/en/tg013.pdf.-   World Seed, Guidelines for the Handling of a Dispute on Essential    Derivation in Lettuce-   Bertier, L. D., et. al., “High-Resolution Analysis of the    Efficiency, Heritability, and Editing Outcomes of    CRISPR/Cas9-Induced Modifications of NCED4 in Lettuce (Lactuca    sativa),” G3: Genes, Genomes, Genetics, 2018, vol. 8, pp. 1513-1521.-   Brotman, Y., et. al., “Resistance Gene Homologues in Melon are    Linked to Genetic Loci Conferring Disease and Pest Resistance”,    Theor Appl Genet, 2002, vol. 104, pp. 1055-1063, DOI    10.1007/s00122-001-0808-x-   Gonai, T., et al., “Abscisic Acid in the Thermoinhibition of Lettuce    Seed Germination and Enhancement of its Catabolism by Gibberellin”,    Journal of Experimental Botany, 2004, vol. 55(394), pp. 111-118.-   Hunter, P. J., et. al., “Oxidative Discolouration in Whole-head and    Cut Lettuce: Biochemical and Environmental Influences on a Complex    Phenotype and Potential Breeding Strategies to Improve Shelf-life,”    Euphytica, 2017, vol. 213(180), DOI 10.1007/s10681-017-1964-7.-   Needleman, S. B., et. al., “A General Method Applicable to the    Search for Similarities in the Amino Acid Sequence of Two Proteins”,    Journal of Molecular Biology, 1970, vol. 48(3), pp. 443-53.-   Nikolova, V., et. al., “Diploidization of Cucumber (Cucumis sativus    L.) Haploids by Colchini Treatment”, Acta Societas Botanicorum    Poloniae, 1996, vol. 65, pp. 311-317.-   Smith, et. al., “Leaf Lettuce Production in California,” 2011,    University of California Agricultural and Natural Resources,    Publication 7216, pp. 1-6.-   Songstad, D. D., et. al., “Genome Editing of Plants,” Critical    Reviews in Plant Sciences, vol. 36, no. 1, pp 1-23.-   Teng, W., et al., “Rapid Regeneration of Lettuce from Suspension    Culture”, HortScience, 1992, vol. 27(9), pp. 1030-1032.-   Teng, W., et al., “Regenerating Lettuce from Suspension Culture in a    2-Liter Bioreactor”, HortScience, 1993, vol. 28(6), pp. 669-671.-   Turini, et. al., “Iceberg Lettuce Production in California,” 2011,    University of California Agricultural and Natural Resources,    Publication 7215, pp. 1-6.-   Vos, P., et al., “AFLP: A New Technique for DNA Fingerprinting”,    Nucleic Acids Research, 1995, vol. 23(21), pp. 4407-4414.-   Wijnker, E., et al., “Hybrid Recreation by Reverse breeding in    Arabidopsis thaliana”, Nature Protocols, 2014, vol. 9, pp. 761-772.    DOI: doi: 10.1038/nprot.2014.049-   Zhang, X., et al., “Genotypic Effects on Tissue Culture Response of    Lettuce Cotyledons”, Journal of Genetics and Breeding, 1992, vol.    46, pp. 287-290.-   US 2008/0222949-   EP 1 197 137 A1-   US 2015/0126380-   WO 2017/144669-   WO 2008/092505-   U.S. Pat. No. 8,237,019    Development of Lettuce Variety NUN 09161 LTL

The inbred variety NUN 09161 LTL was developed from an initial crossbetween lettuce lines. The female and male parents were crossed toproduce seeds. After the cross, progeny was self-pollinated orbackcrossed, followed by pedigree selection and line selection. Lettucevariety NUN 09161 LTL can be propagated by seeds or vegetatively, or byregeneration of a tissue culture. The seeds of lettuce variety NUN 09161LTL can be grown to produce inbred plants and parts thereof (e.g.,lettuce heads and leaves).

The Applicant concluded that lettuce variety NUN 09161 LTL is uniformand stable. This has been established through evaluation ofhorticultural characteristics. Several seed production events resultedin no observable deviation in genetic stability. Lettuce variety NUN09161 LTL has been assigned an inventory ID of 212470303.

DEPOSIT INFORMATION

A total of 2500 seeds of variety NUN 09161 LTL have been depositedaccording to the Budapest Treaty by Nunhems B.V. on Jul. 15, 2022 at theNCIMB Ltd., Ferguson Building, Craibstone Estate, Bucksburn, AberdeenAB21 9YA, United Kingdom (NCIMB). The deposit has been assigned NCIMB43998. A statement indicating the viability of the sample has beenprovided. A deposit of lettuce variety NUN 09161 LTL is also maintainedat Nunhems B.V. The seed lot number for lettuce variety NUN 09161 LTL is30185301002.

The deposit will be maintained in NCIMB for a period of 30 years, or 5years after the most recent request, or for the enforceable life of thepatent whichever is longer and will be replaced if it ever becomesnonviable during that period. Access to the deposits will be availableduring the pendency of this application to persons determined by theDirector of the U.S. Patent Office to be entitled thereto upon request.Subject to 37 C.F.R. § 1.808(b), all restrictions imposed by thedepositor on the availability to the public of the deposited materialwill be irrevocably removed upon the granting of the patent. Applicantdoes not waive any rights granted under this patent on this applicationor under the Plant Variety Protection Act (7 U.S.C. § 2321 et seq.).Accordingly, the requirements of 37 CFR § 1.801-1.809 have beensatisfied.

Characteristics of Lettuce Variety NUN 09161 LTL

The most similar variety to NUN 09161 LTL is referred to as NUN 09060LTL, a variety from Nunhems B.V. with the commercial name Multigreen 60.

In Table 1, the characteristics of lettuce variety NUN 09161 LTL areshown based on trials conducted in Salinas, Calif., USA between March toSeptember 2019. For numerical characteristics, averages were calculated.For non-numerical characteristics, the type/degree were determined.

In Tables 2 and 3, a comparison between lettuce variety NUN 09161 LTLand Reference Variety is shown based on a trial in the USA. Triallocation: Holtville, Calif., USA; Seeding date: Nov. 9, 2019; Harvestingdate: Jan. 30, 2020. In Table 4, the distinguishing characteristicsbetween lettuce variety NUN 09161 LTL and the Reference Variety areshown.

One replication of 30 plants of each variety, from which at least 15plants or plant parts were randomly selected and were used to measurecharacteristics. For numerical characteristics, averages werecalculated. For non-numerical characteristics, the type/degree weredetermined. Similarity and differences between two different plant linesor varieties can be determined by comparing the number of morphologicaland/or physiological characteristics that are the same (i.e.,statistically not significantly different) or that are different (i.e.,statistically significantly different) between the two plant lines orvarieties when grown under the same environmental conditions. Anumerical characteristic is considered to be “the same” when the valuefor a numeric characteristic is not significantly different at the 1%(p<0.01) or 5% (p<0.05) significance level, using T-test, a standardmethod known to the skilled person. A non-numerical characteristics isconsidered to be “the same” when the values have the same “degree” or“type” when scored using USDA and/or UPOV descriptors, for plants grownunder the same environmental conditions. In one aspect, a statisticalanalysis of the quantitative characteristics showing the degree ofsignificance at 5% is provided (see, Tables 5-11).

In another aspect, the disclosure provides a plant having all of thephysiological and morphological characteristics of lettuce variety NUN09161 LTL as presented in Table 1-3, when grown under the sameenvironmental conditions.

TABLE 1 Characteristics of Lettuce Variety NUN 09161 LTL based onSalinas, USA trials between March to September 2019 Application VarietyCharacteristics (NUN 09161 LTL) Vigor/Size: small volume; normal; largevolume Normal Color: light green, normal green, darker green Normalgreen Closed head: open habit even at mature stage; Open habit normal;very closed Head shape: not market acceptable; acceptable, Acceptablevery nice shape Head health: big damage or infections; normal; No damageno damage or infections Frame: lack of frame, normal, good frame NormalBase shape: very veiny or shoulders, open base; Normal and acceptablebase normal and acceptable base; closed base, not shoulders and flatBase health: very sensitive for Botrytis cinerea Strong for basediseases (Grey Mold), Sclerotinia minor and Sclerotinia sclerotiorum(Lettuce Drop), Rhizoctonia solani (Bottom Rot); normal; strong for basediseases Compactness: loose open; normal; very compact Loose openExternal tip burn: tip burn; normal; no tip burn No tip burn Internaltip burn: tip burn and rotting inside the No tip burn head; normal; notip burn Bolting resistance: bolting fast; normal; strong Normal forbolting Taste: bitter; normal; sweet Normal

TABLE 2 Characteristics of Lettuce Variety NUN 09161 LTL and theReference Variety (Exhibit C-USDA descriptors) based on Holtville,California, USA Trial, November 2019-January 2020 Application VarietyReference Variety Characteristics (NUN 09161 LTL) (NUN 09060 LTL) UPOVMorphology: Seed: Seed color: Black Black white, yellow, brown, blackPlant: Degree of overlapping of upper part of leaves: Absent or weakAbsent or weak absent or weak, medium, strong Number of leaves (only forvarieties with degree Few Few of overlapping of upper part of leavesabsent or weak): very few, very few to few, few, few to medium, medium,medium to many, many, many to very many, very many Mature Leaves: Leafattitude: Semi-erect Semi-erect erect, erect to semi-erect, semi-erect,semi-erect to horizontal, horizontal Number of divisions: Medium Manyabsent or very few, very few to few, few, few to medium, medium, mediumto many, many, many to very many, very many Longitudinal section: FlatFlat concave, concave to flat, flat, flat to convex, convex Anthocyanincoloration: Absent or very weak Absent or very weak absent or very weak,very weak to weak, weak, weak to medium, medium, medium to strong,strong, strong to very strong, very strong Color: Green Green green,yellowish green, greyish green Intensity of green color: Medium Mediumvery light, very light to light, light, light to medium, medium, mediumto dark, dark, dark to very dark, very dark Glossiness of upper side:Strong Weak absent or very weak, very weak to weak, weak, weak tomedium, medium, medium to strong, strong, strong to very strong, verystrong Thickness: Thin Thin very thin, thin, medium, thick, very thickBlistering: Absent or slight Absent or slight absent or very weak, veryweak to weak, weak, weak to medium, medium, medium to strong, strong,strong to very strong, very strong Undulation of margin: Strong Mediumto strong absent or very weak, very weak to weak, weak, weak to medium,medium, medium to strong, strong, strong to very strong, very strongType of incisions of margin: Tridentate Tridentate crenate, regularlydentate, irregularly dentate, bidentate, tridentate Depth of incisionsof margin: Medium Medium to deep absent or very shallow, very shallow toshallow, shallow, shallow to medium, medium, medium to deep, deep, deepto very deep, very deep Depth of secondary incisions of margin (onlyShallow Shallow for varieties with type of incisions of marginirregularly dentate, bi- or tri-dentate): very shallow, very shallow toshallow, shallow, shallow to medium, medium, medium to deep, deep, deepto very deep, very deep Density of incisions of margin: Dense dense verysparse, very sparse to sparse, sparse, sparse to medium, medium, mediumto dense, dense, dense to very dense, very dense Venation:Semi-flabellate Semi-flabellate not flabellate, semi-flabellate,flabellate Resistance: Resistance to Bremia lactucae Isolate Bl: 16EUPresent Present Resistance to Bremia lactucae Isolate Bl: 17EU PresentPresent Resistance to Bremia lactucae Isolate Bl: 18EU Present PresentResistance to Bremia lactucae Isolate Bl: 20EU Present PresentResistance to Bremia lactucae Isolate Bl: 21EU Present PresentResistance to Bremia lactucae Isolate Bl: 22EU Present PresentResistance to Bremia lactucae Isolate Bl: 23EU Present PresentResistance to Bremia lactucae Isolate Bl: 24EU Present PresentResistance to Bremia lactucae Isolate Bl: 25EU Present PresentResistance to Bremia lactucae Isolate Bl: 26EU Present PresentResistance to Bremia lactucae Isolate Bl: 27EU Present PresentResistance to Bremia lactucae Isolate Bl: 28EU Present PresentResistance to Bremia lactucae Isolate Bl: 29EU Present PresentResistance to Bremia lactucae Isolate Bl: 30EU Present PresentResistance to Bremia lactucae Isolate Bl: 31EU Present PresentResistance to Bremia lactucae Isolate Bl: 32EU Present PresentResistance to Bremia lactucae Isolate Bl: 33EU Present PresentResistance to Bremia lactucae Isolate Bl: 34EU Present PresentResistance to Bremia lactucae Isolate Bl: 35EU Present PresentResistance to Bremia lactucae Isolate Bl: 36EU Present PresentResistance to Nasonovia ribisnigri (Nr) Biotype Present Present Nr: 0Resistance to Fusarium oxysporum sp. lactucae Absent Present Race 1Resistance to Lettuce Mosaic Virus (LMV) Absent Present Pathotype IIResistance to Lettuce Root Aphids Absent Absent Resistance to Rhizomonassubefaciens (Corky Absent Absent Root) Resistance to Lettuce NecroticStunt Virus Absent Absent (LNSV) USDA Morphology: Plant type:Butterhead, Novita Type, Iceberg Type, Batavia Leaf Leaf Type, Friseed'Amerique Type, Lollo Type, Oakloaf Type, Multi-divided Type, FrilliceType, Cos or Romaine, Bibb/Gem, Stem, Cutting/Whole Leaf, Latin, Other(specify) Spread of frame leaves, cm: 19.70 cm 17.99 cm Butt: Midrib:Flattened Flattened flattened, moderately raised, prominently raised

TABLE 3 Characteristics of Lettuce Variety NUN 09161 LTL and theReference Variety (Non-USDA descriptors) based on Holtville, California,USA Trial, November 2019-January 2020 Application Variety ReferenceVariety Characteristics (NUN 09161 LTL) (NUN 09060 LTL) Plant: Height,cm: 10.22 cm 11.41 cm Weight, g: 99.65 g 94.25 g Head formation: Openhead Open head no head, open head, closed head Diameter: Medium Mediumsmall, medium, large, very large Firmness: Loose Loose Core: Diameter atbase of head, mm (core width): 11.99 mm 10.34 mm Height from base ofhead to apex (mm), average 15.03 mm 12.72 mm (core length): Height frombase of head to apex (mm), range 12.49-17.06 mm 9.86-16.33 mm (min-max):Mature leaves (harvest mature outer leaves): Length, mm: 114.60 mm 114.0mm Width, mm: 115.54 mm 114.68 mm Leaf size: Medium Medium small,medium, large Incision of margin on apical part: Present Present Hue ofgreen color of outer leaves: Yellowish Absent Leaf attitude at 10-12leaf stage: Semi-erect Semi-erect erect, semi-erect, horizontal Leafblade division at 10-12 leaf stage: Divided Divided entire, lobed,divided

TABLE 4 Distinguishing Characteristics between Lettuce Variety NUN 09161LTL and the Reference Variety Application Variety Reference VarietyCharacteristics (NUN 09161 LTL) (NUN 09060 LTL) Plant: Spread of frameleaves, cm: 19.70 cm 17.99 cm Height, cm: 10.22 cm 11.41 cm Core:Diameter at base of head, mm (core width): 11.99 mm 10.34 mm Height frombase of head to apex (mm), 15.03 mm 12.72 mm average (core length):Mature leaves: Number of divisions: Medium Many absent or very few, veryfew to few, few, few to medium, medium, medium to many, many, many tovery many, very many Glossiness of upper side: Strong Weak absent orvery weak, very weak to weak, weak, weak to medium, medium, medium tostrong, strong, strong to very strong, very strong Undulation of margin:Strong Medium to strong absent or very weak, very weak to weak, weak,weak to medium, medium, medium to strong, strong, strong to very strong,very strong Depth of incisions of margin: Medium Medium to deep absentor very shallow, very shallow to shallow, shallow, shallow to medium,medium, medium to deep, deep, deep to very deep, very deep Hue of greencolor of outer leaves: Yellowish Absent

The results of the T-Test show significant differences at 5%significance level between lettuce variety NUN 09161 LTL and theReference Variety for plant spread of frame leaves, plant height, heightfrom base of head to apex (core length), and diameter at based of head(core width) as shown in Tables 5-8.

Table 5 shows a significant difference between lettuce variety NUN 09161LTL and the Reference Variety (p=0.001) for plant spread of frame leaves(cm) based on the results of the trial conducted in the US during thetrial season 2019.

TABLE 5 Application Variety Reference Variety Statistical Parameter (NUN09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 17.30 14.50 Max22.10 21.10 Median 19.45 18.30 Mean 19.70 17.99 Standard deviation 1.371.51

Table 6 shows a significant difference between lettuce variety NUN 09161LTL and the Reference Variety (p=0.004) for plant height (cm) based onthe results of the trial conducted in the US during the trial season2019.

TABLE 6 Application Variety Reference Variety Statistical Parameter (NUN09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 7.80 9.20 Max13.0 13.30 Median 10.20 11.75 Mean 10.22 11.41 Standard deviation 1.191.26

Table 7 shows a significant difference between lettuce variety NUN 09161LTL and the Reference Variety (p<0.001) for height from base of head toapex (core length, mm) based on the results of the trial conducted inthe US during the trial season 2019.

TABLE 7 Application Variety Reference Variety Statistical Parameter (NUN09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 12.49 9.86 Max17.06 16.33 Median 15.17 12.56 Mean 15.03 12.72 Standard deviation 1.151.82

Table 8 shows a significant difference between lettuce variety NUN 09161LTL and the Reference Variety (p<0.001) for diameter at based of head(core width, mm) based on the results of the trial conducted in the USduring the trial season 2019.

TABLE 8 Application Variety Reference Variety Statistical Parameter (NUN09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 9.53 8.78 Max14.23 13.09 Median 12.02 10.12 Mean 11.99 10.34 Standard deviation 1.151.06

The results of the T-Test show no significant differences at 5%significance level between lettuce variety NUN 09161 LTL and theReference Variety for plant weight, mature leaf length, and mature leafwidth as shown in Tables 9-11.

Table 9 shows no significant difference between lettuce variety NUN09161 LTL and the Reference Variety (p=0.284) for plant weight (g) basedon the results of the trial conducted in the US during the trial season2019.

TABLE 9 Application Variety Reference Variety Statistical Parameter (NUN09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 66.0 82.0 Max140.0 132.0 Median 98.0 94.50 Mean 99.65 94.25 Standard deviation 19.3110.82

Table 10 shows no significant difference between lettuce variety NUN09161 LTL and the Reference Variety (p=0.864) for mature leaf length(mm) based on the results of the trial conducted in the US during thetrial season 2019.

TABLE 10 Application Variety Reference Variety Statistical Parameter(NUN 09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 102.3395.62 Max 138.65 137.48 Median 110.71 114.82 Mean 114.60 114.0 Standarddeviation 10.90 10.98

Table 11 shows no significant difference between lettuce variety NUN09161 LTL and the Reference Variety (p=0.721) for mature leaf width (mm)based on the results of the trial conducted in the US during the trialseason 2019.

TABLE 11 Application Variety Reference Variety Statistical Parameter(NUN 09161 LTL) (NUN 09060 LTL) Number of samples 20 20 Min. 107.10105.09 Max 125.37 132.02 Median 116.06 111.51 Mean 115.54 114.68Standard deviation 4.91 9.37

The invention claimed is:
 1. A plant, plant part, or seed of lettucevariety NUN 09161 LTL, wherein a representative sample of seed of saidvariety has been deposited under Accession Number NCIMB
 43998. 2. Theplant part of claim 1, wherein the plant part is a head, a leaf, pollen,an ovule, a fruit, a scion, a rootstock, a cutting, a flower, or a cell.3. A seed that produces the plant of claim
 1. 4. A seed grown on theplant of claim
 1. 5. A lettuce plant grown from the seed of claim
 4. 6.A lettuce plant or a part thereof having all the physiological andmorphological characteristics of the plant of claim
 1. 7. A lettuceplant or a part thereof which does not differ from the plant of claim 1,when determined at the 5% significance level for numericalcharacteristics and determined by type or degree for non-numericalcharacteristics, when grown under the same environmental conditions andwherein a representative sample of seed of lettuce variety NUN 09161 LTLhas been deposited under Accession Number NCIMB
 43998. 8. A tissue orcell culture of regenerable cells of the plant or plant part of claim 1.9. The tissue or cell culture according to claim 8, comprising cells orprotoplasts derived from a plant part suitable for vegetativereproduction, wherein the plant part is an embryo, a meristem, acotyledon, a hypocotyl, pollen, a leaf, an anther, a root, a root tip, apistil, a petiole, a flower, a fruit, a seed, a stem, or a stalk.
 10. Alettuce plant regenerated from the tissue or cell culture of claim 8,wherein the plant has all of the physiological and morphologicalcharacteristics of the plant of lettuce variety NUN 09161 LTL, whendetermined at the 5% significance level for numerical characteristicsand determined by type or degree for non-numerical characteristics forplants grown under the same environmental conditions, and wherein arepresentative sample of seed of said lettuce variety has been depositedunder Accession Number NCIMB
 43998. 11. A method of producing of theplant of claim 1, or a part thereof, said method comprising vegetativepropagation of the plant of lettuce variety NUN 09161 LTL, wherein arepresentative sample of seed of said lettuce variety has been depositedunder Accession Number NCIMB
 43998. 12. The method of claim 11, whereinsaid vegetative propagation comprises regenerating a whole plant from apart of lettuce variety NUN 09161 LTL, wherein a representative sampleof seed of said lettuce variety has been deposited under AccessionNumber NCIMB
 43998. 13. The method of claim 11, wherein said part is acutting, a cell culture, or a tissue culture.
 14. A vegetativelypropagated plant of or part thereof produced by the method of claim 11,wherein the plant has all of the physiological and morphologicalcharacteristics of lettuce variety NUN 09161 LTL, when determined at the5% significance level for numerical characteristics and determined bytype or degree for non-numerical characteristics for plants grown underthe same environmental conditions.
 15. A method of producing a lettuceplant, said method comprising crossing the plant of claim 1 with itselfor a second lettuce plant at least once, and selecting a progeny lettuceplant from said crossing and allowing the progeny lettuce plant to formseed.
 16. A first generation progeny of the lettuce plant of claim 1obtained by crossing the plant of claim 1 with itself or another lettuceplant, wherein the progeny lettuce plant has all of the physiologicaland morphological characteristics of lettuce variety NUN 09161 LTL,wherein a representative sample of seed of said lettuce variety has beendeposited under Accession Number NCIMB
 43998. 17. A method of producinga F1 hybrid lettuce seed, said method comprising crossing a first parentlettuce plant with a second parent lettuce plant and harvesting theresultant hybrid lettuce seed, wherein at the first parent lettuce plantor second parent lettuce plant is the plant of claim
 1. 18. A F1 hybridlettuce seed produced by the method of claim
 17. 19. An F1 hybridlettuce plant or part thereof, produced by growing the seed of claim 18.20. A food, a feed, or a processed product comprising the plant part ofclaim
 2. 21. A container comprising the plant, plant part, or seed ofclaim
 1. 22. A method of introducing a desired trait into the plant ofclaim 1, said method comprises transforming the plant of claim 1 with atransgene that confers the desired trait, wherein the desired trait isyield, storage properties, color, enhanced nutritional quality, malesterility, herbicide tolerance, insect resistance, pest resistance,disease resistance, environmental stress tolerance, modifiedcarbohydrate metabolism, or modified protein metabolism or the mutationoccurs in any of the following genes: Ferulate-5-hydrxylase, dmr1, dmr6,NCED4, PAL, PPO.
 23. A lettuce plant produced by the method of claim 22,wherein the transformed plant contains the desired trait and otherwiseretains all of the morphological and physiological characteristics oflettuce variety NUN 09161 LTL.
 24. A method of introducing a singlelocus conversion into the plant of claim 1, comprising: a. crossing theplant of claim 1 with a second lettuce plant comprising a desired singlelocus to produce F1 progeny plants; b. selecting F1 progeny plants thathave the single locus to produce selected F1 progeny plants; c. crossingthe selected F1 progeny plants with lettuce variety NUN 09161 LTL toproduce backcross progeny plants; d. selecting backcross progeny plantsthat have the single locus and otherwise comprise all of thephysiological and morphological characteristics of lettuce variety NUN09161 LTL to produce selected backcross progeny plants; and e. repeatingsteps (c) and (d) one or more times in succession to produce selectedsecond or higher backcross progeny plants that comprise the single locusand otherwise comprise all of the physiological and morphologicalcharacteristics of lettuce variety NUN 09161 LTL, wherein arepresentative sample of seed of said variety has been deposited underAccession Number NCIMB
 43998. 25. The method of claim 24, wherein thesingle locus confers yield, storage properties, color, enhancednutritional quality, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, or modified proteinmetabolism.
 26. A lettuce plant produced by the method of claim 24,wherein the single locus converted plant comprises the single locusconversion and otherwise has all of the morphological and physiologicalcharacteristics of lettuce variety NUN 09161 LTL.
 27. A method ofproducing a modified lettuce plant, said method comprising mutating atarget gene in lettuce plant or plant part of lettuce variety NUN 09161LTL, wherein a representative sample of seed of said variety has beendeposited under Accession Number NCIMB
 43998. 28. The method of claim27, wherein the target gene modifies a desired trait and wherein thedesired trait is yield, storage properties, color, male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, environmental stress tolerance, modified carbohydratemetabolism, or modified protein metabolism or the mutation occurs in anyof the following genes: Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL,PPO.
 29. The method of claim 27, wherein the target gene is mutated bytargeted gene editing.
 30. A modified lettuce plant produced by themethod of claim 27, wherein the plant contains the desired trait andotherwise all of the physiological and morphological characteristics oflettuce variety NUN 09161 LTL.
 31. A method of producing doubled haploidcells of the plant of claim 1, said method comprising making doubledhaploid cells from haploid cells from the plant or plant part of lettucevariety NUN 09161 LTL, wherein a representative sample of seed of saidlettuce variety has been deposited under Accession Number NCIMB 43998.32. A method of producing a lettuce head or a lettuce leaf, said methodcomprising growing the plant of claim 1 until it develops at least aleaf or a head, and collecting the leaf or head.
 33. A lettuce head or alettuce leaf produced by the method of claim
 32. 34. A method fordetermining the genotype of the plant of claim 1, said method comprisingobtaining a sample of nucleic acids from said plant and detecting insaid nucleic acids a plurality of polymorphisms, thereby determining thegenotype of the plant and storing the results of detecting the pluralityof polymorphisms on a computer readable medium.
 35. A method fordeveloping a lettuce plant in a lettuce breeding program, said methodcomprising applying plant breeding techniques comprising recurrentselection, backcrossing, pedigree breeding, mass selection, mutationbreeding, genetic marker enhanced selection, or genetic transformationto the plant of claim 1 or part thereof.
 36. A method of producing alettuce plant derived from the plant of claim 1, comprising: a.preparing a progeny lettuce plant derived from lettuce variety NUN 09161LTL by crossing the plant of claim 1 with itself or with a secondlettuce plant; b. crossing the progeny plant with itself or a secondlettuce plant to produce seed of a progeny plant of a subsequentgeneration; c. growing a progeny plant of a subsequent generation fromsaid seed and crossing the progeny plant of a subsequent generation withitself or a second lettuce plant; and d. repeating step (b) and/or (c)for at least one more generation to produce a lettuce plant derived fromlettuce variety NUN 09161 LTL.